bioinformatics
TRANSCRIPT
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BIOINFORMATICS
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Contents Overview of Bioinformatics
Bioinformatics
Path to the Bioinformatics
History of Bioinformatics
Ways of modeling Bioinformatic approaches
Static
Dynamic
Structural Bioinformatics
Major Research Areas
Sequence Analysis
Sequence Alignment
Profile Comparison
Sequence Assembly
Gene Prediction
Protein Structure Prediction
Genome Annotation
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Computational Evolutionary Biology
Literature Analysis
Analysis of Gene Expression
Analysis of Regulation
Analysis of Protein Expression
Analysis of Mutations in Cancer
Comparative Genomics
Modeling Biological Systems
High-throughput Image Analysis
Structural Bioinformatic Approaches
Application of Bioinformatics in Various Fields
Molecular medicine
More drug targets
Personalised medicine
Preventative medicine
Gene therapy
Microbial genome applications
Waste cleanup
Climate change
Alternative energy sources
Biotechnology
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Antibiotic resistance
Forensic analysis of microbes
The reality of bioweapon creation
Evolutionary studies
Agriculture
Crops
Insect resistance
Improve nutritional quality
Grow crops in poorer soils and that are drought
resistant
Animals
Comparative studies
Biological Databases
Types of BioInformatics Tools
Homology and Similarity Tools
Protein Function Analysis
Structural Analysis
Sequence Analysis
Bioinformatics Tools
BLAST
FASTA
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EMBOSS
Clustalw
RASMOL
CHEMSKETCH
WINCOOT
AUTODOCK
SWISS PDB VIEWER
Application Programs
JAVA in Bioinformatics
Perl in Bioinformatics
XML in Bioinformatics
C in Bioinformatics
C++ in Bioinformatics
Python in Bioinformatics
R in Bioinformatics
MySQL in Bioinformatics
SQL in Bioinformatics
CUDA in Bioinformatics
MATLAB in Bioinformatics
Microsoft Excel in Bioinformatics
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Bioinformatics Projects
BioJava
BioPerl
BioXML
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BIOINFORMATICS Bioinformatics is a combination of laptop or computer technologies and biology to
manage the immense biological data, primarily genetic data which has been produced
over the years. It entails developing, managing and updating the databases which are
warehouse of the biological data as well as the development of the tools utilized for
analyzing and utilizing this information. The details gathered are then used for the
discovery and development of gene based drugs.
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Overview of Bioinformatics
Introduction
Biology is in the middle of a major paradigm shift driven by computing technology.
Although it is already an informational science in many respects, the field has been
rapidly becoming much more computational and analytical. Rapid progress in genetics
and biochemistry research combined with the tools provided by modern biotechnology
has generated massive volumes of genetic and protein sequence data.
Bioinformatics has been defined as a means for analysing, comparing, graphically
displaying, modeling, storing, systemising, searching, and ultimately distributing
biological information, which includes sequences, structures, function, and phylogeny.
Thus bioinformatics may be defined as a discipline that generates computational tools,
databases, and methods to support genomic and postgenomic research. It comprises the
study of DNA structure and function, gene and protein expression, protein production,
structure and function, genetic regulatory systems, and clinical applications.
Bioinformatics needs the expertise from Computer Science, Mathematics, Statistics,
Medicine, and Biology.
Knowledge Base in Biology
In the last 10 years or so, numerous innovations have seen light and the consequence is
the development of a new biological research paradigm, one that is information-heavy
and computer-driven. As the genetic information is being made as computerized
databases and their sizes are steadily growing, molecular biologists need effective and
efficient computational tools to store and retrieve the cognate information such as
bibliographic or biological information from the databases, to analyze the sequence
patterns they contain and to extract the biological knowledge the sequences have. On the
other hand, there is a strong need for mathematical methods and computational
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techniques for challenging computational tasks such as predicting the three-dimensional
structure of the molecules the sequences represent, and to construct evolutionary trees
from the sequence data. These tools will also be used to learn basic facts about biology
such which sequences of DNA are used to code proteins , which other combinations of
DNA are not used for protein synthesis, for greater understanding of gens and how they
influence diseases.
Biology employs a digital language for represening its information using the four basic
alphabets (A, C, G, T). All the chromosomes in an organism' cell have been represented
and being identified using these alphabets. The demanding challenge here is to determine
how this digital language of the chromosomes is being converted into the three-
dimensional and sometimes four-dimensional languages of living and breathing
organisms.
Information Technology in Biology
As it was found that performing all these above-mentioned tasks manually is nearly
impossible due to the massive volumes of biological data and the preciseness of works, it
became mandatory to use computers for these purposes. Thus this subject of
bioinformatics deals with designing and deploying efficient software tools for
accomplishing the above quoted tasks in a fast and precise manner. So, bridging the gap
between the real world of biology and precise logical nature of computers requires an
interdisciplinary perspective.
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Software and Hardware Advancements in
Biology
The tools of computer science, statistics, and mathematics are very critical for studying
biology as an informational science subject.
Some of the recent advances happened include improved DNA sequencing methods,
new approaches to identify protein structure, and revolutionary methods to monitor the
expression of many genes in parallel. The design of techniques able to deal with different
sources of incomplete and noisy data has become another crucial goal for the
bioinformatics community. In addition, there is the need to implement computational
solutions based on theoretical frameworks to allow scientists to perform complex
inferences about the phenomena under study.
Genomics in the recent past has triggered the development of high-throughput
instrumentation for DNA sequencing, DNA arrays, genotyping, proteomics, etc. These
instruments have catalyzed a new type of science for biology termed discovery science.
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Human Genome Project - An Introduction
The Human Genome Project has encouraged a series of paradigm changes to the view
that biology is an informational science. The draft of the human genome has given us a
genetics parts list of what is necessary for building a human: approximately 35,000
genes, their regulatory regions, a lexicon of motifs that are the building block
components of proteins and genes, and access to the human variability that make us each
different from one user.
Genomes - Discovering Methodology and Study
Discovery science defines all of the elements in a biological system. For example,
sequence of the genome, identification and quantitation of all of the mRNAs or proteins
in a particular cell type - respectively, genome, transcriptome, and the proteome.
Discovery science creates databases of information, in contrast to the more classical
hypothesis-driven science that formulates hypotheses and attempts to test them. The high-
throughput tools both provide the means for discovery science and can assay how global
information sets, for example, transcriptomes or protemes change as systems are
perturbed.
The genomes of the model organisms yeast, worm, fly etc., have demonstrated the
fundamental conservation among all living organisms of the basic informational
pathways. Hence systems can be perturbed in model organisms to gain insight into their
functioning, and these data will provide fundamental insights into human biology. From
the genome, the information pathways and networks can be extracted to begin
understanding their logic of life. Furthermore, different genomes can be compared to
identify similarities and differences in the strategies for the logic of life and these provide
fundamental insights into development, physiology and evolution. The first eukaryotic
genome that has been fully sequenced and annotated is Saccharomyces cerevisiae. This
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highly helps to develop biological and computational tools for genomic and postgenomic
research.
In the era of automated DNA sequencing and revolutionary advances in DNA
sequence analysis, the attention of many researchers is now shifting away from the study
of single genes or small gene clusters to whole genome analyses. Knowing the complete
sequence of a genome is only the first step in understanding how the myriad of
information contained within the genes is transcribed and ultimately translated into
functional proteins. In the post genomic era, functional genomics and proteomic studies
help to obtain an image of the dynamic cell.
System Biology
Biology is a highly informational science. There are mainly two types of biological
information.
1. The information of genes or proteins, which are the molecular machines of life.
2. The information of the regularity networks that coordinate and specify the
expression patterns of the genes and proteins.
All biological information is hierarchical. Initially DNA will change over to mRNA,
which in turn goes to protein. Proteins enact protein interactions, which creates some
informational pathways. These pathways form informational networks, which in turn
become cells. Now cells form networks of cells. Finally an individual is a collection of
cells. A host of individuals forms population and a variety of populations becomes
ecologies. This evolution brings a primary challenge for researchers and scientists to
create tools and mechanisms to capture and integrate these different levels of biological
information and integrate it towards gaining insight of their curious functionings.
All of these paradigm shift lead to the view that the major challenges for biology and
medicine in this new century will be the study of complex systems and the approach
necessary for studying these biological complexities. Here comes a viable approach.
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1. Identify all elements, such as sequence of genomes in the system with currently
available discovery tools
2. Use current knowledge of the sytem to formulate a model predicting its behavior
3. Perturb the system in a model organism using biological, genetic or environmental
perturbations, capture information at all relevant levels, such as DNA, mRNA,
protein, protein interactions, etc. and integrate the collected information
4. Compare theoretical predictions and experimental data, carry out additional
perturbations to bring theory and experiment into closer apposition, integrate new
data into model,
Iterate steps iii) and iv) till the mathematical model can predict the structure of the system
and its systems or emergent properties given particular perturbations.
System Biology - Challenges Ahead
The Integration of technology, biology, and computation.
The integration of the various levels of biological information and the modeling .
The proper annotation of biological information and its its storage and integration
in databases.
The inclusion of other molecules, large and small, in the systems approach.
The integration imperatives of systems biology presents many challenges to
industry and academia.
Conclusion
With the confluence of biology and computer science, the computer applications of
molecular biology are drawing a greater attention among the life science researchers and
scientists these days. As it becomes imperative for biologists to seek the help of
information technology professionals to accomplish the ever growing computational
requirements of a host of exciting and needy biological problems, the synergy between
modern biology and computer science is to blossom in the days to come. Thus the
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research scope for all the mathematical techniques and algorithms coupled with software
programming languages, software development and deployment tools are to get a real
boost. In addition, information technologies such as databases, middleware, graphical
user interface (GUI) design, distributed object computing, storage area networks (SAN),
data compression, network and communication and remote management are all set to
play a very critical role in taking forward the goals for which the bioinformatics field
came into existence.
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Bioinformatics Bioinformatics is the application of computer science and information technology to
the field of biology and medicine. Bioinformatics deals with algorithms, databases and
information systems, web technologies, artificial intelligence and soft computing,
information and computation theory, structural biology, software engineering, data
mining, image processing, modeling and simulation, signal processing, discrete
mathematics, control and system theory, circuit theory, and statistics. Bioinformatics
generates new knowledge as well as the computational tools to create that knowledge.
Bioinformatics:
The use and development of mathematical algorithms and computer programs to obtain insight into biological and medical systems.
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The National Center for Biotechnology Information
(NCBI 2001) defines bioinformatics as:
"Bioinformatics is the field of science in which biology, computer science, and
information technology merges into a single discipline. There are three important sub-
disciplines within bioinformatics: the development of new algorithms and statistics with
which to assess relationships among members of large data sets; the analysis and
interpretation of various types of data including nucleotide and amino acid sequences,
protein domains, and protein structures; and the development and implementation of tools
that enable efficient access and management of different types of information."
From Webopedia:
The application of computer technology to the management of biological information.
Specifically, it is the science of developing computer databases and algorithms to
facilitate and expedite biological research. Bioinformatics is being used largely in the
field of human genome research by the Human Genome Project that has been
determining the sequence of the entire human genome (about 3 billion base pairs) and is
essential in using genomic information to understand diseases. It is also used largely for
the identification of new molecular targets for drug discovery.
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The three terms bioinformatics, computational biology and bioinformation
infrastructure are often times used interchangeably. These three may be defined as
follows:
Bioinformatics refers to database-like activities, involving persistent sets of
data that are maintained in a consistent state over essentially indefinite periods of
time;
Computational biology encompasses the use of algorithmic tools to
facilitate biological analyses; while
Bioinformation infrastructure comprises the entire collective of
information management systems, analysis tools and communication networks
supporting biology. Thus, the latter may be viewed as a computational scaffold of
the former two.
Path to the Bioinformatics
First Learn Biology.
Decide and pick a problem that interests you for experiment.
Find and learn about the Bioinformatics tools.
Learn the Computer Programming Languages.
Experiment on your computer and learn different programming techniques.
The computer has become an essential tool for the biologist just like the microscope.
Eventually the Bioinformatics will become an integral part of the biology.
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History of Bioinformatics
The Modern bioinformatics is can be classified into two broad categories, Biological
Science and computational Science. Here is the data of historical events for both biology
and computer science.
The history of biology in general, B.C. and before the discovery of genetic inheritance
by G. Mendel in 1865, is extremely sketch and inaccurate. This was the start of
Bioinformatics history. Gregor Mendel is known as the "Father of Genetics". He did
experiment on the cross-fertilization of different colors of the same species. He carefully
recorded the data and analyzed the data. Mendel illustrated that the inheritance of traits
could be more easily explained if it was controlled by factors passed down from
generation to generation.
The understanding of genetics has advanced remarkably in the last thirty years. In
1972, Paul berg made the first recombinant DNA molecule using ligase. In that same
year, Stanley Cohen, Annie Chang and Herbert Boyer produced the first recombinant
DNA organism. In 1973, two important things happened in the field of genomics. The
advancement of computing in 1960-70s resulted in the basic methodology of
bioinformatics. However, it is the 1990s when the INTERNET arrived when the full
fledged bioinformatics field was born.
Here are some of the major events in bioinformatics over the last several decades. The
events listed in the list occurred long before the term, "bioinformatics", was coined.
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Bioinformatics Events
1665
Robert Hooke published Micrographia, described the cellular
structure of cork. He also described microscopic examinations
of fossilized plants and animals, comparing their microscopic
structure to that of the living organisms they resembled. He
argued for an organic origin of fossils, and suggested a
plausible mechanism for their formation.
1683 Antoni van Leeuwenhoek discovered bacteria.
1686
John Ray, John Ray's in his book "Historia Plantarum"
catalogued and described 18,600 kinds of plants. His book
gave the first definition of species based upon common
descent.
1843 Richard Owen elaborated the distinction of homology and
analogy.
1864 Ernst Haeckel (Häckel) outlined the essential elements of
modern zoological classification.
1865 Gregory Mendel (1823-1884), Austria, established the theory
of genetic inheritance.
1902 The chromosome theory of heredity is proposed by Sutton
and Boveri, working independently.
1905 The word "genetics" is coined by William Bateson.
1913 First ever linkage map created by Columbia undergraduate
Alfred Sturtevant (working with T.H. Morgan).
1930
Tiselius, Uppsala University, Sweden, A new technique,
electrophoresis, is introduced by Tiselius for separating
proteins in solution. "The moving-boundary method of
studying the electrophoresis of proteins" (published in Nova
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Acta Regiae Societatis Scientiarum Upsaliensis, Ser. IV, Vol. 7,
No. 4)
1946 Genetic material can be transferred laterally between
bacterial cells, as shown by Lederberg and Tatum.
1952
Alfred Day Hershey and Martha Chase proved that the DNA
alone carries genetic information. This was proved on the
basis of their bacteriophage research.
1961 Sidney Brenner, François Jacob, Matthew Meselson, identify
messenger RNA,
1962 Pauling's theory of molecular evolution
1965 Margaret Dayhoff's Atlas of Protein Sequences
1970 Needleman-Wunsch algorithm
1977 DNA sequencing and software to analyze it (Staden)
1981 Smith-Waterman algorithm developed
1981 The concept of a sequence motif (Doolittle)
1982 GenBank Release 3 made public
1982 Phage lambda genome sequenced
1983 Sequence database searching algorithm (Wilbur-Lipman)
1985 FASTP/FASTN: fast sequence similarity searching
1988 National Center for Biotechnology Information (NCBI) created
at NIH/NLM
1988 EMBnet network for database distribution
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1990 BLAST: fast sequence similarity searching
1991 EST: expressed sequence tag sequencing
1993 Sanger Centre, Hinxton, UK
1994 EMBL European Bioinformatics Institute, Hinxton, UK
1995 First bacterial genomes completely sequenced
1996 Yeast genome completely sequenced
1997 PSI-BLAST
1998 Worm (multicellular) genome completely sequenced
1999 Fly genome completely sequenced
2000
Jeong H, Tombor B, Albert R, Oltvai ZN, Barabasi AL. The large-
scale organization of metabolic networks. Nature 2000 Oct
5;407(6804):651-4, PubMed
2000 The genome for Pseudomonas aeruginosa (6.3 Mbp) is
published.
2000 The A. thaliana genome (100 Mb) is secquenced.
2001 The human genome (3 Giga base pairs) is published.
Description
Bioinformatics was applied in the creation and maintenance of a database to store
biological information at the beginning of the "genomic revolution", such as nucleotide
sequences and amino acid sequences. Development of this type of database involved not
only design issues but the development of complex interfaces whereby researchers could
access existing data as well as submit new or revised data.
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In order to study how normal cellular activities are altered in different disease states,
the biological data must be combined to form a comprehensive picture of these activities.
Therefore, the field of bioinformatics has evolved such that the most pressing task now
involves the analysis and interpretation of various types of data. This includes nucleotide
and amino acid sequences, protein domains, and protein structures. The actual process of
analyzing and interpreting data is referred to as computational biology. Important sub-
disciplines within bioinformatics and computational biology include:
The development and implementation of tools that enable efficient access to, and
use and management of, various types of information.
The development of new algorithms (mathematical formulas) and statistics with
which to assess relationships among members of large data sets. For example,
methods to locate a gene within a sequence, predict protein structure and/or
function, and cluster protein sequences into families of related sequences.
The primary goal of bioinformatics is to increase the understanding of biological
processes. What sets it apart from other approaches, however, is its focus on developing
and applying computationally intensive techniques to achieve this goal. Examples
include:
Pattern recognition
Data mining
Machine learning algorithms
Visualization
Major research efforts in the field include
Sequence alignment
Gene finding
Genome assembly
Drug design
Drug discovery
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Protein structure alignment
Protein structure prediction
Prediction of gene expression
Protein–protein interactions
Genome-wide association studies
Modeling of evolution
Interestingly, the term bioinformatics was coined before the "genomic revolution".
Paulien Hogeweg and Ben Hesper introduced the term in 1978 to refer to "the study of
information processes in biotic systems". This definition placed bioinformatics as a field
parallel to biophysics or biochemistry (biochemistry is the study of chemical processes in
biological systems). However, its primary use since at least the late 1980s has been to
describe the application of computer science and information sciences to the analysis of
biological data, particularly in those areas of genomics involving large-scale DNA
sequencing.
Bioinformatics now entails the creation and advancement of databases, algorithms,
computational and statistical techniques and theory to solve formal and practical
problems arising from the management and analysis of biological data.
Over the past few decades rapid developments in genomic and other molecular
research technologies and developments in information technologies have combined to
produce a tremendous amount of information related to molecular biology.
Bioinformatics is the name given to these mathematical and computing approaches used
to glean understanding of biological processes.
Common activities in bioinformatics include mapping and analyzing DNA and protein
sequences, aligning different DNA and protein sequences to compare them, and creating
and viewing 3-D models of protein structures.
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Ways of modeling Bioinformatic
approaches
There are two fundamental ways of modelling a Biological system (e.g., living cell) both
coming under Bioinformatic approaches.
1. Static
2. Dynamic
1. Static
Sequences – Proteins, Nucleic acids and Peptides
Structures – Proteins, Nucleic acids, Ligands (including metabolites and drugs)
and Peptides
Interaction data among the above entities including microarray data and Networks
of proteins, metabolites
2. Dynamic
Systems Biology comes under this category including reaction fluxes and variable
concentrations of metabolites
Multi-Agent Based modelling approaches capturing cellular events such as
signalling, transcription and reaction dynamics
A broad sub-category under bioinformatics is structural
bioinformatics.
Structural Bioinformatics
Structural bioinformatics is the branch of bioinformatics which is related to the
analysis and prediction of the three-dimensional structure of biological macromolecules
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such as proteins, RNA, and DNA. It deals with generalizations about macromolecular 3D
structure such as comparisons of overall folds and local motifs, principles of molecular
folding, evolution, and binding interactions, and structure/function relationships, working
both from experimentally solved structures and from computational models. The term
structural has the same meaning as in structural biology, and structural bioinformatics can
be seen as a part of computational structural biology.
Major Research Areas
1. Sequence Analysis
In bioinformatics, the term sequence analysis refers to the process of subjecting a
DNA, RNA or peptide sequence to any of a wide range of analytical methods to
understand its features, function, structure, or evolution. Methodologies used include
sequence alignment, searches against biological databases, and others. Since the
development of methods of high-throughput production of gene and protein sequences,
the rate of addition of new sequences to the databases increased exponentially. Such a
collection of sequences does not, by itself, increase the scientist's understanding of the
biology of organisms. However, comparing these new sequences to those with known
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functions is a key way of understanding the biology of an organism from which the new
sequence comes. Thus, sequence analysis can be used to assign function to genes and
proteins by the study of the similarities between the compared sequences. Nowadays,
there are many tools and techniques that provide the sequence comparisons (sequence
alignment) and analyze the alignment product to understand its biology.
Sequence analysis in molecular biology includes a very wide range of relevant topics:
1. The comparison of sequences in order to find similarity often to infer if they
are related (homologous).
2. Identification of intrinsic features of the sequence such as active sites, post
translational modification sites, gene-structures, reading frames, distributions
of introns and exons and regulatory elements.
3. Identification of sequence differences and variations such as point mutations
and single nucleotide polymorphism (SNP) in order to get the genetic marker.
4. Revealing the evolution and genetic diversity of sequences and organisms.
5. Identification of molecular structure from sequence alone.
In chemistry, sequence analysis comprises techniques used to do determine the
sequence of a polymer formed of several monomers. In molecular biology and genetics,
the same process is called simply "sequencing".
In marketing, sequence analysis is often used in analytical customer relationship
management applications, such as NPTB models.
History
Since the very first sequences of the insulin protein was characterised by Fred Sanger
in 1951 biologists have been trying to use this knowledge to understand the function of
molecules.
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I. Sequence Alignment
In bioinformatics, a sequence alignment is a way of arranging the sequences of DNA,
RNA, or protein to identify regions of similarity that may be a consequence of functional,
structural, or evolutionary relationships between the sequences. Aligned sequences of
nucleotide or amino acid residues are typically represented as rows within a matrix. Gaps
are inserted between the residues so that identical or similar characters are aligned in
successive columns.
A sequence alignment, produced by ClustalW, of two human zinc finger proteins, identified on the left by GenBank accession number.
Key: Single letters: amino acids. Red: small, hydrophobic, aromatic, not Y. Blue: acidic. Magenta: basic. Green: hydroxyl, amine, amide, basic. Gray: others. "*": identical. ":": conserved substitutions (same colour group). ".": semi-conserved substitution (similar shapes).
Sequence alignments are also used for non-biological sequences, such as those present in
natural language or in financial data.
Interpretation
If two sequences in an alignment share a common ancestor, mismatches can be
interpreted as point mutations and gaps as indels (that is, insertion or deletion mutations)
introduced in one or both lineages in the time since they diverged from one another. In
sequence alignments of proteins, the degree of similarity between amino acids occupying
a particular position in the sequence can be interpreted as a rough measure of how
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conserved a particular region or sequence motif is among lineages. The absence of
substitutions, or the presence of only very conservative substitutions (that is, the
substitution of amino acids whose side chains have similar biochemical properties) in a
particular region of the sequence, suggest that this region has structural or functional
importance. Although DNA and RNA nucleotide bases are more similar to each other
than are amino acids, the conservation of base pairs can indicate a similar functional or
structural role.
Alignment methods
Very short or very similar sequences can be aligned by hand. However, most
interesting problems require the alignment of lengthy, highly variable or extremely
numerous sequences that cannot be aligned solely by human effort. Instead, human
knowledge is applied in constructing algorithms to produce high-quality sequence
alignments, and occasionally in adjusting the final results to reflect patterns that are
difficult to represent algorithmically (especially in the case of nucleotide sequences).
Computational approaches to sequence alignment generally fall into two categories:
global alignments and local alignments. Calculating a global alignment is a form of
global optimization that "forces" the alignment to span the entire length of all query
sequences. By contrast, local alignments identify regions of similarity within long
sequences that are often widely divergent overall. Local alignments are often preferable,
but can be more difficult to calculate because of the additional challenge of identifying
the regions of similarity. A variety of computational algorithms have been applied to the
sequence alignment problem, including slow but formally correct methods like dynamic
programming, and efficient, heuristic algorithms or probabilistic methods that do not
guarantee to find best matches designed for large-scale database search.
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Representations
Alignments are commonly represented both graphically and in text format. In almost
all sequence alignment representations, sequences are written in rows arranged so that
aligned residues appear in successive columns. In text formats, aligned columns
containing identical or similar characters are indicated with a system of conservation
symbols. As in the image above, an asterisk or pipe symbol is used to show identity
between two columns; other less common symbols include a colon for conservative
substitutions and a period for semiconservative substitutions. Many sequence
visualization programs also use color to display information about the properties of the
individual sequence elements; in DNA and RNA sequences, this equates to assigning
each nucleotide its own color. In protein alignments, such as the one in the image above,
color is often used to indicate amino acid properties to aid in judging the conservation of
a given amino acid substitution. For multiple sequences the last row in each column is
often the consensus sequence determined by the alignment; the consensus sequence is
also often represented in graphical format with a sequence logo in which the size of each
nucleotide or amino acid letter corresponds to its degree of conservation.
Sequence alignments can be stored in a wide variety of text-based file formats, many
of which were originally developed in conjunction with a specific alignment program or
implementation. Most web-based tools allow a limited number of input and output
formats, such as FASTA format and GenBank format and the output is not easily
editable. Several conversion programs that provide graphical and/or command line
interfaces are available, such as READSEQ and EMBOSS. There are also several
programming packages which provide this conversion functionality, such as BioPerl and
BioRuby.
Global and local alignments
Global alignments, which attempt to align every residue in every sequence, are most
useful when the sequences in the query set are similar and of roughly equal size. (This
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does not mean global alignments cannot end in gaps.) A general global alignment
technique is the Needleman–Wunsch algorithm, which is based on dynamic
programming. Local alignments are more useful for dissimilar sequences that are
suspected to contain regions of similarity or similar sequence motifs within their larger
sequence context. The Smith–Waterman algorithm is a general local alignment method
also based on dynamic programming. With sufficiently similar sequences, there is no
difference between local and global alignments.
Hybrid methods, known as semiglobal or "glocal" (short for global-local) methods,
attempt to find the best possible alignment that includes the start and end of one or the
other sequence. This can be especially useful when the downstream part of one sequence
overlaps with the upstream part of the other sequence. In this case, neither global nor
local alignment is entirely appropriate: a global alignment would attempt to force the
alignment to extend beyond the region of overlap, while a local alignment might not fully
cover the region of overlap.
Fig: Illustration of global and local alignments demonstrating the 'gappy' quality of global alignments that can occur if sequences are insufficiently similar
Pairwise Alignment
Pairwise sequence alignment methods are used to find the best-matching piecewise
(local) or global alignments of two query sequences. Pairwise alignments can only be
used between two sequences at a time, but they are efficient to calculate and are often
used for methods that do not require extreme precision (such as searching a database for
sequences with high similarity to a query). The three primary methods of producing
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pairwise alignments are dot-matrix methods, dynamic programming, and word methods;
however, multiple sequence alignment techniques can also align pairs of sequences.
Although each method has its individual strengths and weaknesses, all three pairwise
methods have difficulty with highly repetitive sequences of low information content -
especially where the number of repetitions differ in the two sequences to be aligned. One
way of quantifying the utility of a given pairwise alignment is the 'maximum unique
match' (MUM), or the longest subsequence that occurs in both query sequence. Longer
MUM sequences typically reflect closer relatedness.
Dot-matrix methods
The dot-matrix approach, which implicitly produces a family of alignments for
individual sequence regions, is qualitative and conceptually simple, though time-
consuming to analyze on a large scale. In the absence of noise, it can be easy to visually
identify certain sequence features—such as insertions, deletions, repeats, or inverted
repeats—from a dot-matrix plot. To construct a dot-matrix plot, the two sequences are
written along the top row and leftmost column of a two-dimensional matrix and a dot is
placed at any point where the characters in the appropriate columns match—this is a
typical recurrence plot. Some implementations vary the size or intensity of the dot
depending on the degree of similarity of the two characters, to accommodate conservative
substitutions. The dot plots of very closely related sequences will appear as a single line
along the matrix's main diagonal.
Problems with dot plots as an information display technique include: noise, lack of
clarity, non-intuitiveness, difficulty extracting match summary statistics and match
positions on the two sequences. There is also much wasted space where the match data is
inherently duplicated across the diagonal and most of the actual area of the plot is taken
up by either empty space or noise, and, finally, dot-plots are limited to two sequences.
None of these limitations apply to Miropeats alignment diagrams but they have their own
particular flaws.
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Dot plots can also be used to assess repetitiveness in a single sequence. A sequence can
be plotted against itself and regions that share significant similarities will appear as lines
off the main diagonal. This effect can occur when a protein consists of multiple similar
structural domains.
Fig: A DNA dot plot of a human zinc finger transcription factor (GenBank ID NM_002383), showing regional self-similarity. The main diagonal represents the sequence's alignment with itself; lines off the main diagonal represent similar or repetitive patterns within the sequence.
This is a typical example of a recurrence plot.
Dynamic programming
The technique of dynamic programming can be applied to produce global alignments
via the Needleman-Wunsch algorithm, and local alignments via the Smith-Waterman
algorithm. In typical usage, protein alignments use a substitution matrix to assign scores
to amino-acid matches or mismatches, and a gap penalty for matching an amino acid in
one sequence to a gap in the other. DNA and RNA alignments may use a scoring matrix,
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but in practice often simply assign a positive match score, a negative mismatch score, and
a negative gap penalty. (In standard dynamic programming, the score of each amino acid
position is independent of the identity of its neighbors, and therefore base stacking effects
are not taken into account. However, it is possible to account for such effects by
modifying the algorithm.) A common extension to standard linear gap costs, is the usage
of two different gap penalties for opening a gap and for extending a gap. Typically the
former is much larger than the latter, e.g. -10 for gap open and -2 for gap extension. Thus,
the number of gaps in an alignment is usually reduced and residues and gaps are kept
together, which typically makes more biological sense. The Gotoh algorithm implements
affine gap costs by using three matrices.
Dynamic programming can be useful in aligning nucleotide to protein sequences, a
task complicated by the need to take into account frameshift mutations (usually insertions
or deletions). The framesearch method produces a series of global or local pairwise
alignments between a query nucleotide sequence and a search set of protein sequences, or
vice versa. Its ability to evaluate frameshifts offset by an arbitrary number of nucleotides
makes the method useful for sequences containing large numbers of indels, which can be
very difficult to align with more efficient heuristic methods. In practice, the method
requires large amounts of computing power or a system whose architecture is specialized
for dynamic programming. The BLAST and EMBOSS suites provide basic tools for
creating translated alignments (though some of these approaches take advantage of side-
effects of sequence searching capabilities of the tools). More general methods are
available from both commercial sources, such as FrameSearch, distributed as part of the
Accelrys GCG package, and Open Source software such as Genewise.
The dynamic programming method is guaranteed to find an optimal alignment given a
particular scoring function; however, identifying a good scoring function is often an
empirical rather than a theoretical matter. Although dynamic programming is extensible
to more than two sequences, it is prohibitively slow for large numbers of or extremely
long sequences.
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Word Methods
Word methods, also known as k-tuple methods, are heuristic methods that are not
guaranteed to find an optimal alignment solution, but are significantly more efficient than
dynamic programming. These methods are especially useful in large-scale database
searches where it is understood that a large proportion of the candidate sequences will
have essentially no significant match with the query sequence. Word methods are best
known for their implementation in the database search tools FASTA and the BLAST
family. Word methods identify a series of short, nonoverlapping subsequences ("words")
in the query sequence that are then matched to candidate database sequences. The relative
positions of the word in the two sequences being compared are subtracted to obtain an
offset; this will indicate a region of alignment if multiple distinct words produce the same
offset. Only if this region is detected do these methods apply more sensitive alignment
criteria; thus, many unnecessary comparisons with sequences of no appreciable similarity
are eliminated.
In the FASTA method, the user defines a value k to use as the word length with which
to search the database. The method is slower but more sensitive at lower values of k,
which are also preferred for searches involving a very short query sequence. The BLAST
family of search methods provides a number of algorithms optimized for particular types
of queries, such as searching for distantly related sequence matches. BLAST was
developed to provide a faster alternative to FASTA without sacrificing much accuracy;
like FASTA, BLAST uses a word search of length k, but evaluates only the most
significant word matches, rather than every word match as does FASTA. Most BLAST
implementations use a fixed default word length that is optimized for the query and
database type, and that is changed only under special circumstances, such as when
searching with repetitive or very short query sequences. Implementations can be found
via a number of web portals, such as EMBL FASTA and NCBI BLAST.
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Multiple sequence alignment
Multiple sequence alignment is an extension of pairwise alignment to incorporate more
than two sequences at a time. Multiple alignment methods try to align all of the
sequences in a given query set. Multiple alignments are often used in identifying
conserved sequence regions across a group of sequences hypothesized to be
evolutionarily related. Such conserved sequence motifs can be used in conjunction with
structural and mechanistic information to locate the catalytic active sites of enzymes.
Alignments are also used to aid in establishing evolutionary relationships by constructing
phylogenetic trees. Multiple sequence alignments are computationally difficult to produce
and most formulations of the problem lead to NP-complete combinatorial optimization
problems. Nevertheless, the utility of these alignments in bioinformatics has led to the
development of a variety of methods suitable for aligning three or more sequences.
Fig: Alignment of 27 avian influenza hemagglutinin protein sequences colored by residue conservation (top) and residue properties (bottom)
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Dynamic Programming
The technique of dynamic programming is theoretically applicable to any number of
sequences; however, because it is computationally expensive in both time and memory, it
is rarely used for more than three or four sequences in its most basic form. This method
requires constructing the n-dimensional equivalent of the sequence matrix formed from
two sequences, where n is the number of sequences in the query. Standard dynamic
programming is first used on all pairs of query sequences and then the "alignment space"
is filled in by considering possible matches or gaps at intermediate positions, eventually
constructing an alignment essentially between each two-sequence alignment. Although
this technique is computationally expensive, its guarantee of a global optimum solution is
useful in cases where only a few sequences need to be aligned accurately. One method
for reducing the computational demands of dynamic programming, which relies on the
"sum of pairs" objective function, has been implemented in the MSA software package.
Progressive Methods
Progressive, hierarchical, or tree methods generate a multiple sequence alignment by
first aligning the most similar sequences and then adding successively less related
sequences or groups to the alignment until the entire query set has been incorporated into
the solution. The initial tree describing the sequence relatedness is based on pairwise
comparisons that may include heuristic pairwise alignment methods similar to FASTA.
Progressive alignment results are dependent on the choice of "most related" sequences
and thus can be sensitive to inaccuracies in the initial pairwise alignments. Most
progressive multiple sequence alignment methods additionally weight the sequences in
the query set according to their relatedness, which reduces the likelihood of making a
poor choice of initial sequences and thus improves alignment accuracy.
Many variations of the Clustal progressive implementation are used for multiple
sequence alignment, phylogenetic tree construction, and as input for protein structure
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prediction. A slower but more accurate variant of the progressive method is known as T-
Coffee.
Iterative Methods
Iterative methods attempt to improve on the heavy dependence on the accuracy of the
initial pairwise alignments, which is the weak point of the progressive methods. Iterative
methods optimize an objective function based on a selected alignment scoring method by
assigning an initial global alignment and then realigning sequence subsets. The realigned
subsets are then themselves aligned to produce the next iteration's multiple sequence
alignment. Various ways of selecting the sequence subgroups and objective function are
reviewed in.
Motif Finding
Motif finding, also known as profile analysis, constructs global multiple sequence
alignments that attempt to align short conserved sequence motifs among the sequences in
the query set. This is usually done by first constructing a general global multiple
sequence alignment, after which the highly conserved regions are isolated and used to
construct a set of profile matrices. The profile matrix for each conserved region is
arranged like a scoring matrix but its frequency counts for each amino acid or nucleotide
at each position are derived from the conserved region's character distribution rather than
from a more general empirical distribution. The profile matrices are then used to search
other sequences for occurrences of the motif they characterize. In cases where the
original data set contained a small number of sequences, or only highly related
sequences, pseudocounts are added to normalize the character distributions represented in
the motif.
Techniques Inspired by Computer Science
A variety of general optimization algorithms commonly used in computer science have
also been applied to the multiple sequence alignment problem. Hidden Markov models
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have been used to produce probability scores for a family of possible multiple sequence
alignments for a given query set; although early HMM-based methods produced
underwhelming performance, later applications have found them especially effective in
detecting remotely related sequences because they are less susceptible to noise created by
conservative or semi conservative substitutions. Genetic algorithms and simulated
annealing have also been used in optimizing multiple sequence alignment scores as
judged by a scoring function like the sum-of-pairs method. More complete details and
software packages can be found in the main article multiple sequence alignment.
II. Profile Comparison
In 1987 Michael Gribskov, Andrew McLachlan and David Eisenberg introduced the
method of profile comparison for identifying distant similarities between proteins. Rather
than using a single sequence, profile methods use a multiple sequence alignment to
encode a profile which contains information about the conservation level of each residue.
These profiles can then be used to search collections of sequences to find sequences that
are related. Profiles are also known as Position Specific Scoring Matrices (PSSMs). In
1993 a probabilistic interpretation of profiles was introduced by David Haussler and
colleagues using hidden Markov models. These models have become known as profile-
HMMs.
In recent years methods have been developed that allow the comparison of profiles
directly to each other. These are known as profile-profile comparison methods.
III. Sequence Assembly
In bioinformatics, sequence assembly refers to aligning and merging fragments of a
much longer DNA sequence in order to reconstruct the original sequence. This is needed
as DNA sequencing technology cannot read whole genomes in one go, but rather reads
small pieces of between 20 and 1000 bases, depending on the technology used. Typically
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the short fragments, called reads, result from shotgun sequencing genomic DNA, or gene
transcript (ESTs).
The problem of sequence assembly can be compared to taking many copies of a book,
passing them all through a shredder, and piecing the text of the book back together just
by looking at the shredded pieces. Besides the obvious difficulty of this task, there are
some extra practical issues: the original may have many repeated paragraphs, and some
shreds may be modified during shredding to have typos. Excerpts from another book may
also be added in, and some shreds may be completely unrecognizable.
Genome Assemblers
The first sequence assemblers began to appear in the late 1980s and early 1990s as
variants of simpler sequence alignment programs to piece together vast quantities of
fragments generated by automated sequencing instruments called DNA sequencers. As
the sequenced organisms grew in size and complexity (from small viruses over plasmids
to bacteria and finally eukaryotes), the assembly programs used in these genome projects
needed to increasingly employ more and more sophisticated strategies to handle:
Terabytes of sequencing data which need processing on computing clusters.
Identical and nearly identical sequences (known as repeats) which can, in the
worst case, increase the time and space complexity of algorithms exponentially.
Errors in the fragments from the sequencing instruments, which can confound
assembly.
Faced with the challenge of assembling the first larger eukaryotic genomes, the fruit
fly Drosophila melanogaster, in 2000 and the human genome just a year later, scientists
developed assemblers like Celera Assembler and Arachne able to handle genomes of
100-300 million base pairs. Subsequent to these efforts, several other groups, mostly at
the major genome sequencing centers, built large-scale assemblers, and an open source
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effort known as AMOS was launched to bring together all the innovations in genome
assembly technology under the open source framework.
EST Assemblers
Expressed Sequence Tag or EST assembly differs from genome assembly in several
ways. The sequences for EST assembly are the transcribed mRNA of a cell and represent
only a subset of the whole genome. At a first glance, underlying algorithmical problems
differ between genome and EST assembly. For instance, genomes often have large
amounts of repetitive sequences, mainly in the inter-genic parts. Since ESTs represent
gene transcripts, they will not contain these repeats. On the other hand, cells tend to have
a certain number of genes that are constantly expressed in very high amounts
(housekeeping genes), which again leads to the problem of similar sequences present in
high amounts in the data set to be assembled.
Furthermore, genes sometimes overlap in the genome (sense-antisense transcription),
and should ideally still be assembled separately. EST assembly is also complicated by
features like (cis-) alternative splicing, trans-splicing, single-nucleotide polymorphism,
recoding, and post-transcriptional modification.
De-novo vs. Mapping Assembly
In sequence assembly, two different types can be distinguished:
De-novo: assembling short reads to create full-length (sometimes novel)
sequences (see de novo transcriptome assembly)
Mapping: assembling reads against an existing backbone sequence, building a
sequence that is similar but not necessarily identical to the backbone sequence
In terms of complexity and time requirements, de-novo assemblies are orders of
magnitude slower and more memory intensive than mapping assemblies. This is mostly
due to the fact that the assembly algorithm needs to compare every read with every other
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read (an operation that has a complexity of O (n²) but can be reduced to O (n log(n)).
Referring to the comparison drawn to shredded books in the introduction: while for
mapping assemblies one would have a very similar book as template (perhaps with the
names of the main characters and a few locations changed), the de-novo assemblies are
more hardcore in a sense as one would not know beforehand whether this would become
a science book, or a novel, or a catalogue etc.
IV. Gene Prediction
Gene prediction or gene finding refers to the process of identifying the regions of
genomic DNA that encode genes. This includes protein-coding genes as well as RNA
genes, but may also include prediction of other functional elements such as regulatory
regions. Gene finding is one of the first and most important steps in understanding the
genome of a species once it has been sequenced. In general the prediction of bacterial
genes is significantly simpler and more accurate than the prediction of genes in
eukaryotic species that usually have complex intron/exon patterns.
In computational biology gene prediction or gene finding refers to the process of
identifying the regions of genomic DNA that encode genes. This includes protein-coding
genes as well as RNA genes, but may also include prediction of other functional elements
such as regulatory regions. Gene finding is one of the first and most important steps in
understanding the genome of a species once it has been sequenced.
In its earliest days, "gene finding" was based on painstaking experimentation on living
cells and organisms. Statistical analysis of the rates of homologous recombination of
several different genes could determine their order on a certain chromosome, and
information from many such experiments could be combined to create a genetic map
specifying the rough location of known genes relative to each other. Today, with
comprehensive genome sequence and powerful computational resources at the disposal of
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the research community, gene finding has been redefined as a largely computational
problem.
Determining that a sequence is functional should be distinguished from determining
the function of the gene or its product. The latter still demands in vivo experimentation
through gene knockout and other assays, although frontiers of bioinformatics research are
making it increasingly possible to predict the function of a gene based on its sequence
alone.
V. Protein Structure Prediction
Protein threading, also known as fold recognition, is a method of protein modeling (i.e.
computational protein structure prediction) which is used to model those proteins which
have the same fold as proteins of known structures, but do not have homologous proteins
with known structure. It differs from the homology modeling method of structure
prediction as it (protein threading) is used for proteins which do not have their
homologous protein structures deposited in the Protein Data Bank (PDB), whereas
homology modeling is used for those proteins which do. Threading works by using
statistical knowledge of the relationship between the structures deposited in the PDB and
the sequence of the protein which one wishes to model.
The prediction is made by "threading" (i.e. placing, aligning) each amino acid in the
target sequence to a position in the template structure, and evaluating how well the target
fits the template. After the best-fit template is selected, the structural model of the
sequence is built based on the alignment with the chosen template. Protein threading is
based on two basic observations: that the number of different folds in nature is fairly
small (approximately 1300); and that 90% of the new structures submitted to the PDB in
the past three years have similar structural folds to ones already in the PDB (according to
the CATH release notes).
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The 3D structures of molecules are of great importance to their functions in nature.
Since structural prediction of large molecules at an atomic level is largely intractable
problem, some biologists introduced ways to predict 3D structure at a primary sequence
level. This includes biochemical or statistical analysis of amino acid residues in local
regions and structural inference from homologs (or other potentially related proteins)
with known 3D structures.
There have been a large number of diverse approaches to solve the structure prediction
problem. In order to determine which methods were most effective a structure prediction
competition was founded called CASP (Critical Assessment of Structure Prediction).
Fig: Target protein structure (3dsm, shown in ribbons), with Calpha backbones (in gray) of 354 predicted models for it submitted in the CASP8 structure-prediction experiment.
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2. Genome Annotation
In the context of genomics, annotation is the process of marking the genes and other
biological features in a DNA sequence. The first genome annotation software system was
designed in 1995 by Dr. Owen White, who was part of the team at The Institute for
Genomic Research that sequenced and analyzed the first genome of a free-living
organism to be decoded, the bacterium Haemophilus influenzae. Dr. White built a
software system to find the genes (places in the DNA sequence that encode a protein), the
transfer RNA, and other features, and to make initial assignments of function to those
genes. Most current genome annotation systems work similarly, but the programs
available for analysis of genomic DNA are constantly changing and improving.
3. Computational Evolutionary
Biology
Evolutionary biology is the study of the origin and descent of species, as well as their
change over time. Informatics has assisted evolutionary biologists in several key ways; it
has enabled researchers to:
Trace the evolution of a large number of organisms by measuring changes in their
DNA, rather than through physical taxonomy or physiological observations alone.
More recently, compare entire genomes, which permits the study of more complex
evolutionary events, such as gene duplication, horizontal gene transfer, and the
prediction of factors important in bacterial speciation.
Build complex computational models of populations to predict the outcome of the
system over time.
Track and share information on an increasingly large number of species and
organisms.
Future work endeavours to reconstruct the now more complex tree of life.
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The area of research within computer science that uses genetic algorithms is sometimes
confused with computational evolutionary biology, but the two areas are not necessarily
related.
4. Literature Analysis
The growth in the number of published literature makes it virtually impossible to read
every paper, resulting in disjointed subfields of research. Literature analysis aims to
employ computational and statistical linguistics to mine this growing library of text
resources. For example:
Abbreviation recognition - identify the long-form and abbreviation of biological
terms.
Named entity recognition - recognizing biological terms such as gene names.
Protein-protein interaction - identify which proteins interact with which proteins
from text.
The area of research draws from statistics and computational linguistics.
5. Analysis of Gene Expression
The expression of many genes can be determined by measuring mRNA levels with
multiple techniques including microarrays, expressed cDNA sequence tag (EST)
sequencing, serial analysis of gene expression (SAGE) tag sequencing, massively parallel
signature sequencing (MPSS), RNA-Seq, also known as "Whole Transcriptome Shotgun
Sequencing" (WTSS), or various applications of multiplexed in-situ hybridization. All of
these techniques are extremely noise-prone and/or subject to bias in the biological
measurement, and a major research area in computational biology involves developing
statistical tools to separate signal from noise in high-throughput gene expression studies.
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Such studies are often used to determine the genes implicated in a disorder: one might
compare microarray data from cancerous epithelial cells to data from non-cancerous cells
to determine the transcripts that are up-regulated and down-regulated in a particular
population of cancer cells.
6. Analysis of Regulation
Regulation is the complex orchestration of events starting with an extracellular signal
such as a hormone and leading to an increase or decrease in the activity of one or more
proteins. Bioinformatics techniques have been applied to explore various steps in this
process. For example, promoter analysis involves the identification and study of sequence
motifs in the DNA surrounding the coding region of a gene. These motifs influence the
extent to which that region is transcribed into mRNA. Expression data can be used to
infer gene regulation: one might compare microarray data from a wide variety of states of
an organism to form hypotheses about the genes involved in each state.
In a single-cell organism, one might compare stages of the cell cycle, along with
various stress conditions (heat shock, starvation, etc.). One can then apply clustering
algorithms to that expression data to determine which genes are co-expressed. For
example, the upstream regions (promoters) of co-expressed genes can be searched for
over-represented regulatory elements.
7. Analysis of Protein Expression
Protein microarrays and high throughput (HT) mass spectrometry (MS) can provide a
snapshot of the proteins present in a biological sample. Bioinformatics is very much
involved in making sense of protein microarray and HT MS data; the former approach
faces similar problems as with microarrays targeted at mRNA, the latter involves the
problem of matching large amounts of mass data against predicted masses from protein
sequence databases, and the complicated statistical analysis of samples where multiple,
but incomplete peptides from each protein are detected.
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8. Analysis of Mutations in Cancer
In cancer, the genomes of affected cells are rearranged in complex or even
unpredictable ways. Massive sequencing efforts are used to identify previously unknown
point mutations in a variety of genes in cancer. Bioinformaticians continue to produce
specialized automated systems to manage the sheer volume of sequence data produced,
and they create new algorithms and software to compare the sequencing results to the
growing collection of human genome sequences and germline polymorphisms. New
physical detection technologies are employed, such as oligonucleotide microarrays to
identify chromosomal gains and losses (called comparative genomic hybridization), and
single-nucleotide polymorphism arrays to detect known point mutations. These detection
methods simultaneously measure several hundred thousand sites throughout the genome,
and when used in high-throughput to measure thousands of samples, generate terabytes of
data per experiment. Again the massive amounts and new types of data generate new
opportunities for bioinformaticians. The data is often found to contain considerable
variability, or noise, and thus Hidden Markov model and change-point analysis methods
are being developed to infer real copy number changes.
Another type of data that requires novel informatics development is the analysis of
lesions found to be recurrent among many tumors.
9. Comparative Genomics
Comparative genomics is the study of the relationship of genome structure and
function across different biological species or strains. Comparative genomics is an
attempt to take advantage of the information provided by the signatures of selection to
understand the function and evolutionary processes that act on genomes. While it is still a
young field, it holds great promise to yield insights into many aspects of the evolution of
modern species. The sheer amount of information contained in modern genomes (3.2
gigabases in the case of humans) necessitates that the methods of comparative genomics
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are automated. Gene finding is an important application of comparative genomics, as is
discovery of new, non-coding functional elements of the genome.
Comparative genomics exploits both similarities and differences in the proteins, RNA,
and regulatory regions of different organisms to infer how selection has acted upon these
elements. Those elements that are responsible for similarities between different species
should be conserved through time (stabilizing selection), while those elements
responsible for differences among species should be divergent (positive selection).
Finally, those elements that are unimportant to the evolutionary success of the organism
will be unconserved (selection is neutral).
One of the important goals of the field is the identification of the mechanisms of
eukaryotic genome evolution. It is however often complicated by the multiplicity of
events that have taken place throughout the history of individual lineages, leaving only
distorted and superimposed traces in the genome of each living organism. For this reason
comparative genomics studies of small model organisms (for example the model
Caenorhabditis elegans and closely related Caenorhabditis briggsae) are of great
importance to advance our understanding of general mechanisms of evolution.
Having come a long way from its initial use of finding functional proteins, comparative
genomics is now concentrating on finding regulatory regions and siRNA molecules.
Recently, it has been discovered that distantly related species often share long conserved
stretches of DNA that do not appear to code for any protein (see conserved non-coding
sequence). One such ultra-conserved region, that was stable from chicken to chimp has
undergone a sudden burst of change in the human lineage, and is found to be active in the
developing brain of the human embryo.
Computational approaches to genome comparison have recently become a common
research topic in computer science. A public collection of case studies and
demonstrations is growing, ranging from whole genome comparisons to gene expression
analysis. This has increased the introduction of different ideas, including concepts from
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systems and control, information theory, strings analysis and data mining. It is anticipated
that computational approaches will become and remain a standard topic for research and
teaching, while multiple courses will begin training students to be fluent in both topics.
Fig: Human FOXP2 gene and evolutionary conservation is shown in and multiple alignment (at bottom of figure) in this image from the UCSC Genome Browser. Note that conservation tends
to cluster around coding regions (exons).
10. Modeling Biological Systems
Systems biology involves the use of computer simulations of cellular subsystems (such
as the networks of metabolites and enzymes which comprise metabolism, signal
transduction pathways and gene regulatory networks) to both analyze and visualize the
complex connections of these cellular processes. Artificial life or virtual evolution
attempts to understand evolutionary processes via the computer simulation of simple
(artificial) life forms.
Modelling biological systems is a significant task of systems biology and mathematical
biology. Computational systems biology aims to develop and use efficient algorithms,
data structures, visualization and communication tools with the goal of computer
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modelling of biological systems. It involves the use of computer simulations of biological
systems, like cellular subsystems (such as the networks of metabolites and enzymes
which comprise metabolism, signal transduction pathways and gene regulatory networks)
to both analyze and visualize the complex connections of these cellular processes.
Artificial life or virtual evolution attempts to understand evolutionary processes via the
computer simulation of simple (artificial) life forms.
Overview
It is understood that an unexpected emergent property of a complex system is a result
of the interplay of the cause-and-effect among simpler, integrated parts. Biological
systems manifest many important examples of emergent properties in the complex
interplay of components. Traditional study of biological systems requires reductive
methods in which quantities of data are gathered by category, such as concentration over
time in response to a certain stimulus. Computers are critical to analysis and modelling of
these data. The goal is to create accurate real-time models of a system's response to
environmental and internal stimuli, such as a model of a cancer cell in order to find
weaknesses in its signalling pathways, or modelling of ion channel mutations to see
effects on cardiomyocytes and in turn, the function of a beating heart.
A monograph on this topic summarizes an extensive amount of published research in
this area up to 1987, including subsections in the following areas: computer modelling in
biology and medicine, arterial system models, neuron models, biochemical and
oscillation networks, quantum automata, quantum computers in molecular biology and
genetics, cancer modelling, neural nets, genetic networks, abstract relational biology,
metabolic-replication systems, category theory applications in biology and medicine,
automata theory, cellular automata, tessallation models and complete self-reproduction,
chaotic systems in organisms, relational biology and organismic theories. This published
report also includes 390 references to peer-reviewed articles by a large number of
authors.
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Standards
By far the most widely accepted standard format for storing and exchanging models in
the field is the Systems Biology Markup Language (SBML) The SBML.org website
includes a guide to many important software packages used in computational systems
biology. Other markup languages with different emphases include BioPAX and CellML.
Particular Tasks
Cellular model
Creating a cellular model has been a particularly challenging task of systems biology
and mathematical biology. It involves the use of computer simulations of the many
cellular subsystems such as the networks of metabolites and enzymes which comprise
metabolism, signal transduction pathways and gene regulatory networks to both analyze
and visualize the complex connections of these cellular processes.
The complex network of biochemical reaction/transport processes and their spatial
organization make the development of a predictive model of a living cell a grand
challenge for the 21st century.
In 2006, the National Science Foundation (NSF) put forward a grand challenge for
systems biology in the 21st century to build a mathematical model of the whole cell. E-
Cell Project aims "to make precise whole cell simulation at the molecular level possible".
CytoSolve developed by V. A. Shiva Ayyadurai and C. Forbes Dewey, Jr. of Department
of Biological Engineering at the Massachusetts Institute of Technology, provided a
method to model the whole cell by dynamically integrating multiple molecular pathway
models.
A dynamic computer model of intracellular signaling was the basis for Merrimack
Pharmaceuticals to discover the target for their cancer medicine MM-111.
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Membrane computing is the task of modelling specifically a cell membrane.
Fig: Signal transduction pathways
Protein Folding
Protein structure prediction is the prediction of the three-dimensional structure of a
protein from its amino acid sequence — that is, the prediction of its secondary, tertiary,
and quaternary structure from its primary structure. Structure prediction is fundamentally
different from the inverse problem of protein design. Protein structure prediction is one of
the most important goals pursued by bioinformatics and theoretical chemistry; it is highly
important in medicine (for example, in drug design) and biotechnology (for example, in
the design of novel enzymes). Every two years, the performance of current methods is
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assessed in the CASP experiment (Critical Assessment of Techniques for Protein
Structure Prediction).
Human Biological Systems
Brain Model
The Blue Brain Project is an attempt to create a synthetic brain by reverse-engineering
the mammalian brain down to the molecular level. The aim of the project, founded in
May 2005 by the Brain and Mind Institute of the École Polytechnique in Lausanne,
Switzerland, is to study the brain's architectural and functional principles. The project is
headed by the Institute's director, Henry Markram. Using a Blue Gene supercomputer
running Michael Hines's NEURON software, the simulation does not consist simply of an
artificial neural network, but involves a partially biologically realistic model of neurons.
It is hoped by its proponents that it will eventually shed light on the nature of
consciousness.
There are a number of sub-projects, including the Cajal Blue Brain, coordinated by the
Supercomputing and Visualization Center of Madrid (CeSViMa), and others run by
universities and independent laboratories in the UK, U.S., and Israel. The Human Brain
Project builds on the work of the Blue Brain Project. It is one of six pilot projects in the
Future Emerging Technologies Research Program of the European Commission,
competing for a billion euro funding.
Model of the Immune System
The last decade has seen the emergence of a growing number of simulations of the
immune system.
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Virtual Liver
The Virtual Liver project is a 43 million euro research program funded by the German
Government, made up of seventy research group distributed across Germany. The goal is
to produce a virtual liver, a dynamic mathematical model that represents human liver
physiology, morphology and function.
11. High-throughput Image Analysis
Computational technologies are used to accelerate or fully automate the processing,
quantification and analysis of large amounts of high-information-content biomedical
imagery. Modern image analysis systems augment an observer's ability to make
measurements from a large or complex set of images, by improving accuracy, objectivity,
or speed. A fully developed analysis system may completely replace the observer.
Although these systems are not unique to biomedical imagery, biomedical imaging is
becoming more important for both diagnostics and research. Some examples are:
High-throughput and high-fidelity quantification and sub-cellular localization
(high-content screening, cytohistopathology, Bioimage informatics)
Morphometrics
Clinical image analysis and visualization
Determining the real-time air-flow patterns in breathing lungs of living animals
Quantifying occlusion size in real-time imagery from the development of and
recovery during arterial injury
Making behavioral observations from extended video recordings of laboratory
animals
Infrared measurements for metabolic activity determination
Inferring clone overlaps in DNA mapping, e.g. the Sulston score
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12. Structural Bioinformatic Approaches
Prediction of Protein Structure
Protein structure prediction is another important application of bioinformatics. The
amino acid sequence of a protein, the so-called primary structure, can be easily
determined from the sequence on the gene that codes for it. In the vast majority of cases,
this primary structure uniquely determines a structure in its native environment.
Knowledge of this structure is vital in understanding the function of the protein. For lack
of better terms, structural information is usually classified as one of secondary, tertiary
and quaternary structure. A viable general solution to such predictions remains an open
problem. As of now, most efforts have been directed towards heuristics that work most of
the time.
One of the key ideas in bioinformatics is the notion of homology. In the genomic
branch of bioinformatics, homology is used to predict the function of a gene: if the
sequence of gene A, whose function is known, is homologous to the sequence of gene B,
whose function is unknown, one could infer that B may share A's function. In the
structural branch of bioinformatics, homology is used to determine which parts of a
protein are important in structure formation and interaction with other proteins. In a
technique called homology modeling, this information is used to predict the structure of a
protein once the structure of a homologous protein is known. This currently remains the
only way to predict protein structures reliably.
One example of this is the similar protein homology between hemoglobin in humans
and the hemoglobin in legumes (leghemoglobin). Both serve the same purpose of
transporting oxygen in the organism. Though both of these proteins have completely
different amino acid sequences, their protein structures are virtually identical, which
reflects their near identical purposes.
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Other techniques for predicting protein structure include protein threading and de novo
physics-based modeling.
Molecular Interaction
Protein–protein interaction prediction is a field combining bioinformatics and
structural biology in an attempt to identify and catalog physical interactions between
pairs or groups of proteins. Understanding protein–protein interactions is important for
the investigation of intracellular signaling pathways, modelling of protein complex
structures and for gaining insights into various biochemical processes. Experimentally,
physical interactions between pairs of proteins can be inferred from a variety of
experimental techniques, including yeast two-hybrid systems, protein-fragment
complementation assays (PCA), affinity purification/mass spectrometry, protein
microarrays, fluorescence resonance energy transfer (FRET), and Microscale
Thermophoresis (MST). Efforts to experimentally determine the interactome of numerous
species are ongoing, and a number of computational methods for interaction prediction
have been developed in recent years.
Methods
Proteins that interact are more likely to co-evolve, therefore, it is possible to make
inferences about interactions between pairs of proteins based on their phylogenetic
distances. It has also been observed in some cases that pairs of interacting proteins have
fused orthologues in other organisms. In addition, a number of bound protein complexes
have been structurally solved and can be used to identify the residues that mediate the
interaction so that similar motifs can be located in other organisms.
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Phylogenetic Profiling
Phylogenetic profiling finds pairs of protein families with similar patterns of presence
or absence across large numbers of species. This method identifies pairs likely to act in
the same biological process, but does not necessarily imply physical interaction.
Prediction of Co-evolved Protein Pairs Based on
Similar Phylogenetic Trees
This method involves using a sequence search tool such as BLAST for finding
homologues of a pair of proteins, then building multiple sequence alignments with
alignment tools such as Clustal. From these multiple sequence alignments, phylogenetic
distance matrices are calculated for each protein in the hypothesized interacting pair. If
the matrices are sufficiently similar (as measured by their Pearson correlation coefficient)
they are deemed likely to interact.
Identification of Homologous Interacting Pairs
This method consists of searching whether the two sequences have homologues which
form a complex in a database of known structures of complexes. The identification of the
domains is done by sequence searches against domain databases such as Pfam using
BLAST. If more than one complex of Pfam domains is identified, then the query
sequences are aligned using a hidden Markov tool called HMMER to the closest
identified homologues, whose structures are known. Then the alignments are analysed to
check whether the contact residues of the known complex are conserved in the alignment.
Identification of Structural Patterns
This method builds a library of known protein–protein interfaces from the PDB, where
the interfaces are defined as pairs of polypeptide fragments that are below a threshold
slightly larger than the Van der Waals radius of the atoms involved. The sequences in the
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library are then clustered based on structural alignment and redundant sequences are
eliminated. The residues that have a high (generally >50%) level of frequency for a given
position are considered hotspots. This library is then used to identify potential
interactions between pairs of targets, providing that they have a known structure (i.e.
present in the PDB).
Bayesian Network Modelling
Bayesian methods integrate data from a wide variety of sources, including both
experimental results and prior computational predictions, and use these features to assess
the likelihood that a particular potential protein interaction is a true positive result. These
methods are useful because experimental procedures, particularly the yeast two-hybrid
experiments, are extremely noisy and produce many false positives, while the previously
mentioned computational methods can only provide circumstantial evidence that a
particular pair of proteins might interact.
3D Template-Based Protein Complex Modelling
This method makes use of known protein complex structures to predict as well as
structurally model interactions between query protein sequences. The prediction process
generally starts by employing a sequence based method (e.g. Interolog) to search for
protein complex structures that are homologous to the query sequences. These known
complex structures are then used as templates to structurally model the interaction
between query sequences. This method has the advantage of not only inferring protein
interactions but also suggests models of how proteins interact structurally, which can
provide some insights into the atomic level mechanism of that interaction. On the other
hand, the ability for this method to makes a prediction is limited to a relatively small
number of known protein complex structures.
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Supervised Learning Problem
The problem of PPI prediction can be framed as a supervised learning problem. In this
paradigm the known protein interactions supervise the estimation of a function that can
predict whether an interaction exists or not between two proteins given data about the
proteins (e.g., expression levels of each gene in different experimental conditions,
location information, phylogenetic profile, etc.).
Relationship to Docking Methods
The field of protein–protein interaction prediction is closely related to the field of
protein–protein docking, which attempts to use geometric and steric considerations to fit
two proteins of known structure into a bound complex. This is a useful mode of inquiry
in cases where both proteins in the pair have known structures and are known (or at least
strongly suspected) to interact, but since so many proteins do not have experimentally
determined structures, sequence-based interaction prediction methods are especially
useful in conjunction with experimental studies of an organism's interactome.
Application of Bioinformatics in
Various Fields Bioinformatics is the use of IT in biotechnology for the data storage, data warehousing
and analyzing the DNA sequences. In Bioinfomatics knowledge of many branches are
required like biology, mathematics, computer science, laws of physics & chemistry, and
of course sound knowledge of IT to analyze biotech data. Bioinformatics is not limited to
the computing data, but in reality it can be used to solve many biological problems and
find out how living things works.
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It is the comprehensive application of mathematics (e.g., probability and statistics),
science (e.g., biochemistry), and a core set of problem-solving methods (e.g., computer
algorithms) to the understanding of living systems.
These include the following:
1. Molecular medicine
1.1 More drug targets
1.2 Personalised medicine
1.3 Preventative medicine
1.4 Gene therapy
2. Microbial genome applications
2.1 Waste cleanup
2.2 Climate change
2.3 Alternative energy sources
2.4 Biotechnology
2.5 Antibiotic resistance
2.6 Forensic analysis of microbes
2.7 The reality of bioweapon creation
2.8 Evolutionary studies
3. Agriculture
3.1 Crops
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3.2 Insect resistance
3.3 Improve nutritional quality
3.4 Grow crops in poorer soils and that are drought resistant
4. Animals
5. Comparative studies
1. Molecular Medicine
Molecular medicine is a broad field, where physical, chemical, biological and medical
techniques are used to describe molecular structures and mechanisms, identify
fundamental molecular and genetic errors of disease, and to develop molecular
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interventions to correct them. The molecular medicine perspective emphasizes cellular
and molecular phenomena and interventions rather than the previous conceptual and
observational focus on patients and their organs.
In November 1949, with the seminal paper, "Sickle Cell Anemia, a Molecular
Disease", in Science magazine, Linus Pauling, Harvey Itano and their collaborators laid
the groundwork for establishing the field of molecular medicine. In 1956, Roger J.
Williams wrote Biochemical Individuality, a prescient book about genetics, prevention
and treatment of disease on a molecular basis, and nutrition which is now variously
referred to as individualized medicine and orthomolecular medicine. Another paper in
Science by Pauling in 1968, introduced and defined this view of molecular medicine that
focuses on natural and nutritional substances used for treatment and prevention.
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The human genome will have profound effects on the fields of biomedical research and
clinical medicine. Every disease has a genetic component. This may be inherited (as is
the case with an estimated 3000-4000 hereditary disease including Cystic Fibrosis and
Huntingtons disease) or a result of the body's response to an environmental stress which
causes alterations in the genome (eg. cancers, heart disease, diabetes..).
The completion of the human genome means that we can search for the genes directly
associated with different diseases and begin to understand the molecular basis of these
diseases more clearly. This new knowledge of the molecular mechanisms of disease will
enable better treatments, cures and even preventative tests to be developed.
1.1 More Drug Targets
At present all drugs on the market target only about 500 proteins. With an improved
understanding of disease mechanisms and using computational tools to identify and
validate new drug targets, more specific medicines that act on the cause, not merely the
symptoms, of the disease can be developed. These highly specific drugs promise to have
fewer side effects than many of today's medicines.
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1.2 Personalised Medicine
Personalized medicine is a medical model that proposes the customization of
healthcare, with all decisions and practices being tailored to the individual patient by use
of genetic or other information. Practical application outside of long established
considerations like a patient's family history, social circumstances, environment and
behaviors are very limited so far and practically no progress has been made in the last
decade.
Clinical medicine will become more personalised with the development of the field of
pharmacogenomics. This is the study of how an individual's genetic inheritence affects
the body's response to drugs. At present, some drugs fail to make it to the market because
a small percentage of the clinical patient population show adverse affects to a drug due to
sequence variants in their DNA.
As a result, potentially lifesaving drugs never make it to the marketplace. Today,
doctors have to use trial and error to find the best drug to treat a particular patient as those
with the same clinical symptoms can show a wide range of responses to the same
treatment. In the future, doctors will be able to analyse a patient's genetic profile and
prescribe the best available drug therapy and dosage from the beginning.
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1.3 Preventative Medicine
Preventive medicine or preventive care consists of measures taken to prevent diseases,
(or injuries) rather than curing them or treating their symptoms. This contrasts in method
with curative and palliative medicine, and in scope with public health methods (which
work at the level of population health rather than individual health).
Preventive medicine strategies are typically described as taking place at the primary,
secondary, tertiary and quaternary prevention levels. In addition, the term primal
prevention has been used to describe all measures taken to ensure fetal well-being and
prevent any long-term health consequences from gestational history and/or disease. The
rationale for such efforts is the evidence demonstrating the link between fetal well-being,
or "primal health," and adult health. Primal prevention strategies typically focus on
providing future parents with: education regarding the consequences of epigenetic
influences on their child, sufficient leave time for both parents, and financial support if
required. This includes parenting in infancy as well.
Simple examples of preventive medicine include hand washing, breastfeeding, and
immunizations. Preventive care may include examinations and screening tests tailored to
an individual's age, health, and family history. For example, a person with a family
history of certain cancers or other diseases would begin screening at an earlier age and/or
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more frequently than those with no such family history. On the other side of preventive
medicine, some nonprofit organizations, such as the Northern California Cancer Center,
apply epidemiologic research towards finding ways to prevent diseases.
Prevention levels
Doctor’s side
Disease
absent present
Patient’s
side
Illness
absent
Primary
prevention
illness absent
disease absent
Secondary
prevention
illness absent
disease present
present
Quaternary
prevention
illness present
disease absent
Tertiary
prevention
illness present
disease present
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Definitions
With the specific details of the genetic mechanisms of diseases being unravelled, the
development of diagnostic tests to measure a person’s susceptibility to different diseases
may become a distinct reality. Preventative actions such as change of lifestyle or having
treatment at the earliest possible stages when they are more likely to be successful, could
result in huge advances in our struggle to conquer disease.
1.4 Gene Therapy
Gene therapy is the use of DNA as a pharmaceutical agent to treat disease. It derives
its name from the idea that DNA can be used to supplement or alter genes within an
individual's cells as a therapy to treat disease. The most common form of gene therapy
involves using DNA that encodes a functional, therapeutic gene in order to replace a
mutated gene. Other forms involve directly correcting a mutation, or using DNA that
encodes a therapeutic protein drug (rather than a natural human gene) to provide
treatment. In gene therapy, DNA that encodes a therapeutic protein is packaged within a
"vector", which is used to get the DNA inside cells within the body. Once inside, the
Level Definition
Primary
prevention
Methods to avoid occurrence of disease. Most population-based health
promotion efforts are of this type.
Secondary
prevention
Methods to diagnose and treat existent disease in early stages before it
causes significant morbidity.
Tertiary
prevention
Methods to reduce negative impact of extant disease by restoring function
and reducing disease-related complications.
Quaternary
prevention
Methods to mitigate or avoid results of unnecessary or excessive
interventions in the health system.
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DNA becomes expressed by the cell machinery, resulting in the production of therapeutic
protein, which in turn treats the patient's disease.
Fig: Gene therapy using an Adenovirus vector. A new gene is inserted into an adenovirus. If the treatment is successful, the new gene will make functional protein to treat a disease.
In the not too distant future, the potential for using genes themselves to treat disease
may become a reality. Gene therapy is the approach used to treat, cure or even prevent
disease by changing the expression of a person’s genes. Currently, this field is in its
infantile stage with clinical trials for many different types of cancer and other diseases
ongoing.
1.5 Drug Designing
Drug Discovery process is an expensive and time consuming process that may be
succeed or may not. Retrospective analysis of pharmaceutical industry during the 1990’s
estimate that each new drug in the market takes an average 12-15 years to develop,
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costing in the region of $1.3 billion. In addition, one in nine compounds that enters
clinical trials makes it to the market.
In the present era, the emerging new diseases are very much responsible for the
advancement in the field of Computer Aided Drug Design (CADD). Drug Designing is
the innovative boom in the field of Bioinformatics which is very cost and time effective.
Hence, computers are playing a vital role in the Drug Designing.
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Drug Discovery process have formed by four factors ; Scientific Knowledge, Available
Technology, Human Resources and Diseases. The Scientific Knowledge, Available
Technology, Human Resource provide the strategic assets by which the strategies were
formed.
Once a new compound has been identified in the laboratory, medicines are developed as
follows:
I. PRECLINICAL TESTING:
A pharmaceutical company conducts laboratory and animal studies to show biological
activity of the compound against the target disease, and the compound is evaluated for
safety.
II. INVESTIGATIONAL NEW DRUG APPLICATION (IND):
After completing preclinical testing, a company files an IND with the U.S. Food and
Drug Administration (FDA) to begin to test the drug in people. The IND becomes
effective if FDA does not disapprove it within 30days. The IND shows results of
previous experiments; how, where and by whom the new studies will be conducted; the
chemical structure of the compound; how it is thought to work in the body; any toxic
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effects found in the animal studies; and how the compound is manufactured. All clinical
trials must be reviewed and approved by the Institutional Review Board (IRB) where the
trials will be conducted.
III. CLINICAL TRIALS:
PHASE-1:
In this studies are primarily concerned with assessing the drug candidate’s safety. A
small number of healthy volunteers are given the compound to test what happens to the
drug in the human body – how it is absorbed, metabolized and excreted. About 70% of
drug candidates pass this initial phase of testing.
PHASE-2:
The drug candidate is tested for efficacy. Usually, this is explored in a randomised trial
where the compound or a drug is given to several hundred patients with the condition or
disease to be treated. Depending on the condition the trial can last from several months to
several number of years. The output is an increased understanding of the safety of the
compound and clear information about effectiveness. If a drug passes this phase, then it
can be considered that the drug can be considered truly a drug.
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PHASE-3:
A drug is tested in several hundred to several thousand patients. This provides a more
thorough understanding of the drug’s effectiveness, benefits and the range of possible
adverse reactions. These trials typically last several years.
IV. APPROVAL:
Once FDA approves an NDA, the new medicine becomes available for physicians to
prescribe. A company must continue to submit periodic reports to FDA, including any
cases of adverse reactions and appropriate quality control records.
Hence, in this way a drug is said to be passed through different trials and atlast to the
market.
2. Microbial Genome Applications
Microorganisms are ubiquitous, that is they are found everywhere. They have been
found surviving and thriving in extremes of heat, cold, radiation, salt, acidity and
pressure. They are present in the environment, our bodies, the air, food and water.
Traditionally, use has been made of a variety of microbial properties in the baking,
brewing and food industries. The arrival of the complete genome sequences and their
potential to provide a greater insight into the microbial world and its capacities could
have broad and far reaching implications for environment, health, energy and industrial
applications. For these reasons, in 1994, the US Department of Energy (DOE) initiated
the MGP (Microbial Genome Project) to sequence genomes of bacteria useful in energy
production, environmental cleanup, industrial processing and toxic waste reduction.
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By studying the genetic material of these organisms, scientists can begin to understand
these microbes at a very fundamental level and isolate the genes that give them their
unique abilities to survive under extreme conditions.
2.1 Waste Cleanup
Deinococcus radiodurans is known as the world's toughest bacteria and it is the most
radiation resistant organism known. Scientists are interested in this organism because of
its potential usefulness in cleaning up waste sites that contain radiation and toxic
chemicals.
Microbial Genome Program (MGP) scientists are determining the DNA sequence of
the genome of C. crescentus, one of the organisms responsible for sewage treatment.
2.2 Climate Change
Increasing levels of carbon dioxide emission, mainly through the expanding use of
fossil fuels for energy, are thought to contribute to global climate change. Recently, the
DOE (Department of Energy, USA) launched a program to decrease atmospheric carbon
dioxide levels. One method of doing so is to study the genomes of microbes that use
carbon dioxide as their sole carbon source.
2.3 Alternative Energy Sources
Scientists are studying the genome of the microbe Chlorobium tepidum which has an
unusual capacity for generating energy from light.
2.4 Biotechnology
The archaeon Archaeoglobus fulgidus and the bacterium Thermotoga maritima have
potential for practical applications in industry and government-funded environmental
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remediation. These microorganisms thrive in water temperatures above the boiling point
and therefore may provide the DOE, the Department of Defence, and private companies
with heat-stable enzymes suitable for use in industrial processes.
Other industrially useful microbes include, Corynebacterium glutamicum which is of
high industrial interest as a research object because it is used by the chemical industry for
the biotechnological production of the amino acid lysine. The substance is employed as a
source of protein in animal nutrition. Lysine is one of the essential amino acids in animal
nutrition. Biotechnologically produced lysine is added to feed concentrates as a source of
protein, and is an alternative to soybeans or meat and bonemeal.
Xanthomonas campestris pv. is grown commercially to produce the exopolysaccharide
xanthan gum, which is used as a viscosifying and stabilising agent in many industries.
Lactococcus lactis is one of the most important micro-organisms involved in the dairy
industry, it is a non-pathogenic rod-shaped bacterium that is critical for manufacturing
dairy products like buttermilk, yogurt and cheese. This bacterium, Lactococcus lactis
ssp., is also used to prepare pickled vegetables, beer, wine, some breads and sausages and
other fermented foods. Researchers anticipate that understanding the physiology and
genetic make-up of this bacterium will prove invaluable for food manufacturers as well
as the pharmaceutical industry, which is exploring the capacity of L. lactis to serve as a
vehicle for delivering drugs.
2.5 Antibiotic Resistance
Scientists have been examining the genome of Enterococcus faecalis a leading cause
of bacterial infection among hospital patients. They have discovered a virulence region
made up of a number of antibiotic-resistant genes that may contribute to the bacterium's
transformation from a harmless gut bacteria to a menacing invader. The discovery of the
region, known as a pathogenicity island, could provide useful markers for detecting
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pathogenic strains and help to establish controls to prevent the spread of infection in
wards.
2.6 Forensic Analysis of Microbes
Scientists used their genomic tools to help distinguish between the strain of Bacillus
anthracis that was used in the summer of 2001 terrorist attack in Florida with that of
closely related anthrax strains.
2.7 The Reality of Bioweapon Creation
Scientists have recently built the virus poliomyelitis using entirely artificial means.
They did this using genomic data available on the Internet and materials from a mail-
order chemical supply. The research was financed by the US Department of Defence as
part of a biowarfare response program to prove to the world the reality of bioweapons.
The researchers also hope their work will discourage officials from ever relaxing
programs of immunisation. This project has been met with very mixed feelings.
2.8 Evolutionary Studies
The sequencing of genomes from all three domains of life, eukaryota, bacteria and
archaea means that evolutionary studies can be performed in a quest to determine the tree
of life and the last universal common ancestor.
3. Agriculture
The sequencing of the genomes of plants and animals should have enormous benefits
for the agricultural community. Bioinformatic tools can be used to search for the genes
within these genomes and to elucidate their functions. This specific genetic knowledge
could then be used to produce stronger, more drought, disease and insect resistant crops
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and improve the quality of livestock making them healthier, more disease resistant and
more productive.
3.1 Crops
Comparative genetics of the plant genomes has shown that the organisation of their
genes has remained more conserved over evolutionary time than was previously believed.
These findings suggest that information obtained from the model crop systems can be
used to suggest improvements to other food crops. Arabidopsis thaliana (water cress) and
Oryza sativa (rice) are examples of available complete plant genomes.
3.2 Insect Resistance
Genes from Bacillus thuringiensis that can control a number of serious pests have been
successfully transferred to cotton, maize and potatoes. This new ability of the plants to
resist insect attack means that the amount of insecticides being used can be reduced and
hence the nutritional quality of the crops is increased.
3.3 Improve Nutritional Quality
Scientists have recently succeeded in transferring genes into rice to increase levels of
Vitamin A, iron and other micronutrients. This work could have a profound impact in
reducing occurrences of blindness and anaemia caused by deficiencies in Vitamin A and
iron respectively.
Scientists have inserted a gene from yeast into the tomato, and the result is a plant
whose fruit stays longer on the vine and has an extended shelf life.
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3.4 Grow in Poorer Soils and Drought Resistant
Progress has been made in developing cereal varieties that have a greater tolerance for
soil alkalinity, free aluminium and iron toxicities. These varieties will allow agriculture to
succeed in poorer soil areas, thus adding more land to the global production base.
Research is also in progress to produce crop varieties capable of tolerating reduced water
conditions.
4. Animals
Sequencing projects of many farm animals including cows, pigs and sheep are now
well under way in the hope that a better understanding of the biology of these organisms
will have huge impacts for improving the production and health of livestock and
ultimately have benefits for human nutrition.
5. Comparative Studies
Analysing and comparing the genetic material of different species is an important
method for studying the functions of genes, the mechanisms of inherited diseases and
species evolution. Bioinformatics tools can be used to make comparisons between the
numbers, locations and biochemical functions of genes in different organisms.
Organisms that are suitable for use in experimental research are termed model
organisms. They have a number of properties that make them ideal for research purposes
including short life spans, rapid reproduction, being easy to handle, inexpensive and they
can be manipulated at the genetic level.
An example of a human model organism is the mouse. Mouse and human are very
closely related (>98%) and for the most part we see a one to one correspondence between
genes in the two species. Manipulation of the mouse at the molecular level and genome
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comparisons between the two species can and is revealing detailed information on the
functions of human genes, the evolutionary relationship between the two species and the
molecular mechanisms of many human diseases.
Biological Databases The collection of the biological data on a computer which can be manipulated to appear
in varying arrangements and subsets is regarded as a database. The biological information
can be stored in different databases. Each database has its own website with unique
navigation tools.
Nucleotide sequence databases: The nucleotide sequence data
submitted by the scientists and genome sequencing groups is at the databases
namely GenBank, EMBL (European Molecular Biology Laboratory) and DDBI
(DNA Data Bank of Japan). There is a good coordination between these three
databases as they are synchronized on daily basis.
Protein sequence databases: These are usually prepared from
existing literature and/or in consultation with the experts. In fact, these databases
represent the translated DNA databases.
Molecular structure of databases: The three dimensional (3-D)
structures of macro molecules are determined by X-ray crystallography and
nuclear magnetic resonance (NMR). PDB and SCOP are the primary databases of
biological molecules.
Other databases: KEGG database is an important one that provides
information on the current knowledge of molecular biology and cell biology with
special reference to information on metabolic pathways, interacting molecules and
genes.
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Types of BioInformatics Tools The Bioinformatics tools are the software programs for the saving, retrieving and analysis
of Biological data and extracting the information from them.
Factors that must be taken into consideration when designing these tools are:
The end user (the biologist) may not be a frequent user of computer technology
and thus it should be very user friendly.
These software tools must be made available over the internet given the global
distribution of the scientific research community.
The Bioinformatics Tools may be categorized into following categories:
1. Homology and Similarity Tools
2. Protein Function Analysis
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3. Structural Analysis
4. Sequence Analysis
1.Homology and Similarity Tools
The term homology implies a common evolutionary relationship between two traits -
whether they are DNA sequences or bristle patterns on a fly's nose. Homologous
sequences are sequences that are related by divergence from a common ancestor. Thus
the degree of similarity between two sequences can be measured while their homology is
a case of being either true of false. This set of tools can be used to identify similarities
between novel query sequences of unknown structure and function and database
sequences whose structure and function have been elucidated.
2. Protein Function Analysis
Function Analysis is Identification and mapping of all functional elements (both
coding and non-coding) in a genome. This group of programs allow you to compare your
protein sequence to the secondary (or derived) protein databases that contain information
on motifs, signatures and protein domains. Highly significant hits against these different
pattern databases allow you to approximate the biochemical function of your query
protein.
3. Structural Analysis
These sets of tools allow you to compare structures with the known structure
databases. The function of a protein is more directly a consequence of its structure rather
than its sequence with structural homologs tending to share functions. The determination
of a protein's 2D/3D structure is crucial in the study of its function.
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4. Sequence Analysis
This set of tools allows you to carry out further, more detailed analysis on your query
sequence including evolutionary analysis, identification of mutations, hydropathy
regions, CpG islands and compositional biases. The identification of these and other
biological properties are all clues that aid the search to elucidate the specific function of
your sequence.
Bioinformatics Tools
1. BLAST
The Basic Local Alignment Search Tool (BLAST) for comparing gene and protein
sequences against others in public databases, now comes in several types including PSI-
BLAST, PHI-BLAST, and BLAST 2 sequences. Specialized BLASTs are also available
for human, microbial, malaria, and other genomes, as well as for vector contamination,
immunoglobulins, and tentative human consensus sequences.
2. FASTA
A database search tool used to compare a nucleotide or peptide sequence to a sequence
database. The program is based on the rapid sequence algorithm described by Lipman
and Pearson. It was the first widely used algorithm for database similarity searching. The
program looks for optimal local alignments by scanning the sequence for small matches
called "words". Initially, the scores of segments in which there are multiple word hits are
calculated ("init1"). Later the scores of several segments may be summed to generate an
"initn" score. An optimized alignment that includes gaps is shown in the output as "opt".
The sensitivity and speed of the search are inversely related and controlled by the "k-tup"
variable which specifies the size of a "word".
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3. EMBOSS
EMBOSS (The European Molecular Biology Open Software Suite) is a new, free open
source software analysis package specially developed for the needs of the molecular
biology user community. Within EMBOSS you will find around 100 programs
(applications) for sequence alignment, database searching with sequence patterns, protein
motif identification and domain analysis, nucleotide sequence pattern analysis, codon
usage analysis for small genomes, and much more.
4. Clustalw
ClustalW is a general purpose multiple sequence alignment program for DNA or
proteins. It produces biologically meaningful multiple sequence alignments of divergent
sequences, calculates the best match for the selected sequences, and lines them up so that
the identities, similarities and differences can be seen.
5. RASMOL
RasMol is a computer program written for molecular graphics visualization intended
and used primarily for the depiction and exploration of biological macromolecule
structures, such as those found in the Protein Data Bank. It was originally developed by
Roger Sayle in the early 90s.
Historically, it was an important tool for molecular biologists since the extremely
optimized program allowed the software to run on (then) modestly powerful personal
computers. Before RasMol, visualization software ran on graphics workstations that, due
to their expense, were less accessible to scholars. RasMol has become an important
educational tool as well as continuing to be an important tool for research in structural
biology.
Protein Databank (PDB) files can be downloaded for visualization from the Research
Collaboratory for Structural Bioinformatics (RCSB) bank. These have been uploaded by
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researchers who have characterized the structure of molecules usually by X-ray
crystallography or NMR spectroscopy.
6. CHEMSKETCH
ACD/ChemSketch is an advanced chemical drawing tool and is the accepted interface
for the industry's best NMR and molecular property predictions, nomenclature, and
analytical data handling software. ACD/ChemSketch is also available as freeware, with
functionalities that are highly competitive with other popular commercial software
packages. The freeware contains tools for 2D structure cleaning, 3D optimization and
viewing, InChI generation and conversion, drawing of polymers, organometallics, and
Markush structures—capabilities that are not even included in some of the commercial
packages from other software producers. Also included is an IUPAC systematic naming
capability for molecules with fewer than 50 atoms and 3 rings. The capabilities of
ACD/ChemSketch can be further extended and customized by programming.
The commercial version of ACD/ChemSketch offers additional capabilities above and
beyond the freeware offering. It includes a number of advanced features including a
dictionary of more than 165,000 trivial, common, and trade names with their
corresponding structures. It allows the user to view SDfiles, and search Microsoft Word
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or Adobe PDF reports, SDfiles, molfiles, and CambridgeSoft ChemDraw files by
chemical structure, substructure, or structure similarity.
7. WINCOOT
WinCoot is developed by Bernhard Lohkamp & Paul Emsley and is used by 4 users of
Software Informer. The most popular versions of this product among our users are: 0.0,
0.1 and 0.3. The name of the program executable file is cmd.exe.
8. AUTODOCK
AutoDock is a suite of automated docking tools. It is designed to predict how small
molecules, such as substrates or drug candidates, bind to a receptor of known 3D
structure. AutoDock actually consists of two main programs: AutoDock performs the
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docking of the ligand to a set of grids describing the target protein; AutoGrid pre-
calculates these grids. In addition to using them for docking, the atomic affinity grids can
be visualised. This can help, for example, to guide organic synthetic chemists design
better binders. We have also developed a graphical user interface called AutoDockTools,
or ADT for short, which amongst other things helps to set up which bonds will treated as
rotatable in the ligand and to analyze dockings.
AutoDock has applications in:
X-ray crystallography
Structure-based drug design
Lead optimization
Virtual screening (HTS)
Combinatorial library design
Protein-protein docking
Chemical mechanism studies
9. SWISS PDB VIEWER
Swiss-PdbViewer (aka DeepView) has been developped since 1994 by Nicolas Guex.
Swiss-PdbViewer is tightly linked to SWISS-MODEL, an automated homology modeling
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server developed within the Swiss Institute of Bioinformatics (SIB) at the Structural
Bioinformatics Group at the Biozentrum in Basel.
Swiss-PdbViewer is an application that provides a user friendly interface allowing to
analyze several proteins at the same time. The proteins can be superimposed in order to
deduce structural alignments and compare their active sites or any other relevant parts.
Amino acid mutations, H-bonds, angles and distances between atoms are easy to obtain
thanks to the intuitive graphic and menu interface.
Application Programs
1. JAVA in Bioinformatics
Due to Platform independence nature of Java, it is emerging as a key player in
bioinformatics. Physiome Sciences' computer-based biological simulation technologies
and Bioinformatics Solutions' PatternHunter are two examples of the growing adoption of
Java in bioinformatics.
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2. Perl in Bioinformatics
Perl is also being used in the processing of biological data. One example of perl project
is BioPerl project.
3. XML in Bioinformatics
Extensible Markup Language (XML) is a markup language that defines a set of rules
for encoding documents in a format that is both human-readable and machine-readable. It
is defined in the XML 1.0 Specification produced by the W3C, and several other related
specifications, all gratis open standards.
The design goals of XML emphasize simplicity, generality, and usability over the
Internet. It is a textual data format with strong support via Unicode for the languages of
the world. Although the design of XML focuses on documents, it is widely used for the
representation of arbitrary data structures, for example in web services.
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4. C in Bioinformatics
C is a general-purpose programming language initially developed by Dennis Ritchie
between 1969 and 1973 at Bell Labs. Its design provides constructs that map efficiently
to typical machine instructions, and therefore it found lasting use in applications that had
formerly been coded in assembly language—most notably system software like the Unix
operating system. C is one of the most widely used programming languages of all time,
and there are very few computer architectures for which a C compiler does not exist.
5. C++ in Bioinformatics
C++ (pronounced "cee plus plus") is a statically typed, free-form, multi-paradigm,
compiled, general-purpose programming language. It is regarded as an intermediate-level
language, as it comprises a combination of both high-level and low-level language
features. Developed by Bjarne Stroustrup starting in 1979 at Bell Labs, it adds object
oriented features, such as classes, and other enhancements to the C programming
language. Originally named C with Classes, the language was renamed C++ in 1983, as a
pun involving the increment operator.
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6. Python in Bioinformatics
Python is a general-purpose, high-level programming language whose design
philosophy emphasizes code readability. Its syntax is said to be clear and expressive.
Python has a large and comprehensive standard library.
7. R in Bioinformatics
R is an open source programming language and software environment for statistical
computing and graphics. The R language is widely used among statisticians for
developing statistical software and data analysis.
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8. MySQL in Bioinformatics
MySQL is the world's most used open source relational database management system
(RDBMS) that runs as a server providing multi-user access to a number of databases.
9. SQL in Bioinformatics
SQL (sometimes referred to as Structured Query Language) is a special-purpose
programming language designed for managing data in relational database management
systems (RDBMS).
Originally based upon relational algebra and tuple relational calculus, its scope
includes data insert, query, update and delete, schema creation and modification, and data
access control.
10. CUDA in Bioinformatics
Compute Unified Device Architecture (CUDA) is a parallel computing architecture
developed by Nvidia for graphics processing. CUDA is the computing engine in Nvidia
graphics processing units (GPUs) that is accessible to software developers through
variants of industry standard programming languages. Programmers use 'C for CUDA' (C
with Nvidia extensions and certain restrictions), compiled through a PathScale or Open64
C compiler, to code algorithms for execution on the GPU. CUDA architecture shares a
range of computational interfaces with two competitors: the Khronos Group's OpenCL
and Microsoft's DirectCompute. Third party wrappers are also available for Python, Perl,
Fortran, Java, Ruby, Lua, Haskell, MATLAB, IDL, and native support in Mathematica.
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CUDA programming in the web browser is freely available for individual non-
commercial purposes in NCLab.
11. MATLAB in Bioinformatics
MATLAB (matrix laboratory) is a numerical computing environment and fourth-
generation programming language. Developed by MathWorks, MATLAB allows matrix
manipulations, plotting of functions and data, implementation of algorithms, creation of
user interfaces, and interfacing with programs written in other languages, including C,
C++, Java, and Fortran.
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12. Microsoft Excel in Bioinformatics
Microsoft Excel is a commercial spreadsheet application written and distributed by
Microsoft for Microsoft Windows and Mac OS X. It features calculation, graphing tools,
pivot tables, and a macro programming language called Visual Basic for Applications. It
has been a very widely applied spreadsheet for these platforms, especially since version 5
in 1993, and it has almost completely replaced Lotus 1-2-3 as the industry standard for
spreadsheets. Excel forms part of Microsoft Office. The current versions are 2010 for
Microsoft Windows and 2011 for Mac OS X.
Bioinformatics Projects
1. BioJava
The BioJava Project is providing the Java tool for the processing of data in Java.
2. BioPerl
The BioPerl project many module for biological data processing.
3. BioXML
A part of the BioPerl project, this is a resource to gather XML documentation, DTDs and
XML aware tools for biology in one location.