frontiers of biotechnology chapter 9 mr. scott. manipulating dna how can scientists manipulate the...
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Frontiers of Biotechnology
Chapter 9Mr. Scott
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Manipulating DNA• How can
scientists manipulate the code?
• Scientists use their knowledge of the structure of DNA and its chemical properties to study and change DNA molecules.
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Manipulating DNA• Tools of Molecular
Biology– Genetic
Engineering• Making changes in
the DNA code of a living organism
– DNA Extraction• Open a cell
(mechanical and chemical methods)
• Use a chemical to separate the DNA from the rest of the cell parts (an alcohol)
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Manipulating DNA–Cutting DNA
• DNA molecules are very long
• Restriction enzymes
– Enzymes that cuts DNA at a specific sequence of nucleotides
• Restriction Site– Site where
restriction enzyme cuts DNA
• Hundreds of different restriction sites from enzymes
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Manipulating DNA• Sticky ends
– these are the cut ends of the DNA–Pieces can stick to corresponding nucleotides
–Used in biotechnology
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Manipulating DNA– Separating DNA
• Gel electrophoresis– A mixture of
DNA fragments is placed at one end of a porous gel (negative end), and an electric voltage is applied to the gel
–The molecules will move from negative to positive
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Manipulating DNA• Restriction Maps–A pattern of bands showing lengths of cut DNA
–Can be used to study mutations
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Manipulating DNA
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Copying DNA• PCR or Polymerase chain reaction
• Photocopying DNA over and over
• Use strands of DNA that are called primers
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Copying DNA• 3 – Step
Process– Need DNA to
be copied, 4 bases, and two primers
• Primers – short segment of used for starting point
• Steps1. Separating2. Binding3. Copying
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DNA Fingerprinting• DNA
Fingerprinting–DNA that can
be used for identification
–Cut with enzyme and run through gel electrophoresis
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DNA Fingerprinting• Greatest differences in DNA are found in regions not used for genes–Each person has differing numbers of repeating units in similar regions
–Differences are found by gel electrophoresis
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DNA Fingerprinting• Indentification and Probability–Identification depends on probability
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DNA Fingerprinting• Uses of
Fingerprinting–Criminal cases–Family
relations• Paternity• Kinship for
immigration purposes
• Species studies
• Agriculture
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Genetic Engineering• Cloning
– Clone – a genetically identical model• Gene or organism
–Transfer DNA between cells
–Dolly – The cloned sheep from 1997
–Controversial
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Genetic Engineering• Cloning has potential
benefits.–organs for transplant into humans
–save endangered species• Cloning raises concerns.
–low success rate–clones “imperfect” and less healthy than original animal
–decreased biodiversity
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Genetic Engineering• Genetic Engineering–Changing DNA to give it new traits
–Can insert healthy genes or cloned genes
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Manipulating DNA–Cutting and
Pasting• Recombinant DNA
• Enzymes join genes from one strand of DNA with genes from another strand of DNA
• DNA synthesizers
– Enzymes join artificial pieces of DNA with natural pieces
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Genetic Engineering• Transforming
Bacteria–Foreign DNA is
joined to plasmid• Plasmid – small,
circular DNA molecule
• Genetic marker – a gene that makes it possible see which bacteria are carrying the plasmid (that is marked) and which bacteria are not.
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Genetic Engineering• Transformation–When a cell takes in foreign DNA and joins it with the existing DNA
–This external DNA becomes part of the cells DNA
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Genetic Engineering• New genes can
be added to an organism’s DNA.– Genetic
engineering involves changing an organism’s DNA to give it new traits.
– Genetic engineering is based on the use of recombinant DNA.
– Recombinant DNA contains genes from more than one organism.
(bacterial DNA)
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Genetic Engineering• Transgenic
Organisms–Organisms that contain genes from other species
–Firefly and tobacco plant
–Genetic engineering has increased the growth of biotechnology
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Genetic Engineering• Transforming Plant
Cells– Using bacterium
that normally infect plant cells and cause tumors
– Taking away the cell wall some plant cells will take up DNA on their own
– If successful recombinant DNA will be found in a chromosome of the cell
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Genetic Engineering• Transforming
Animal Cells– DNA can be
injected directly into the nucleus of an egg
– Enzymes help insert the new DNA into the eggs existing DNA
– New DNA contains markers that tell which cells picked up the DNA
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Genetic Engineering
–Transgenic microorganisms• Bacteria reproduce quickly
• Uses:–Can grow human
proteins• Insulin, growth
hormone, and clotting factor
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Genetic Engineering
–Transgenic Animals• Mimic human body for testing
• Faster growing animals
• No more food poisoning
• Grow human proteins
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Genetic Engineering• Transgenic
animals are used to study diseases and gene functions.– Transgenic mice
used to study development and disease
– Gene knockout mice used to study gene function• Leptin – helps
control food intake – common for the obese
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Genetic Engineering
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Genetic Engineering–Transgenic Plants
• Natural insecticide
• Resist weed killing chemicals
• Food resistant to spoiling
• Weather resistant plants
• Grow vitamins or proteins
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Genomics and Bioinformatics
• Genomics is the study of genomes.–Can include the sequencing of the genome
–Comparisons of genomes within and across species
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Genomics and Bioinformatics
• DNA Sequencing–Determining
the order of DNA nucleotides in genes or in genomes.
–Genomes of several different organisms have been sequenced
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Genomics and Bioinformatics
• The Human Genome Project– Has sequenced
all of the DNA base pairs of human chromosomes.
– Analyzed DNA from a few people
– Still working to identify and map human genes
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Genomics and Bioinformatics
• Technology and Genes– Bioinformatics
• The use of computer databases to organize and analyze biological data.
– DNA microarrays • Used to study
the expression of many genes at once.
– Proteomics – The study and
comparison of proteins.
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Genetic Screening and Therapy
• Genetic screening can detect genetic disorders.– Determines risk of having
or passing on a geneticdisorder
– Used to detect specificgenes or proteins
– Can detect some genesrelated to an increasedrisk of cancer
– Can detect some genesknown to cause geneticdisorders
DMD N
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Genetic Screening and Therapy
• Gene therapy is the replacement of faulty genes.
• Several experimental techniques are used for gene therapy.–Genetically engineered viruses
used to “infect” a patient’s cells– Insert gene to stimulate
immune system to attack cancer cells
– Insert “suicide” genes into cancer cells that activate a drug
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Genetic Screening and Therapy
• Gene therapy has many technical challenges.–Inserting gene into correct cells
–Controlling gene expression–Determining effect on other genes