the molecular biology of the gene. experiments have shown that dna is the genetic material – dna...
TRANSCRIPT
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THE MOLECULAR BIOLOGY OF THE GENE
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THE MOLECULAR BIOLOGY OF THE GENE
• EXPERIMENTS HAVE SHOWN THAT DNA IS THE GENETIC MATERIAL– DNA HAS BEEN KNOWN ABOUT FOR OVER 100
YEARS; HOWEVER, KNOWLEDGE OF ITS ROLE AS GENETIC MATERIAL IS RELATIVELY NEW
– ORIGINALLY, IT WAS THOUGHT THAT PROTEIN WAS IN CHARGE OF TRAITS (WHY DO YOU THINK THEY BELIEVED PROTEINS WERE THE ANSWER?)
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THE MOLECULAR BIOLOGY OF THE GENE
• GRIFFITH’S EXPERIMENT
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THE MOLECULAR BIOLOGY OF THE GENE
• GRIFFITH’S SETUP– INVOLVED A TYPE OF BACTERIA (STREPTOCOCCUS)– UTILIZED HEAT SHOCK TO DENATURE PROTEINS
• GRIFFITH’S RESULTS– HARMLESS FORM (R FORM) WAS TURNED INTO
HARMFUL FORM (S FORM) WHEN HEAT-SHOCKED “S” WAS MIXED WITH NORMAL “R”
– IDENTIFIED DNA IS GENETIC CARRIER
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THE MOLECULAR BIOLOGY OF THE GENE
• SCIENTISTS WERE RESISTANT TO GRIFFITH’S FINDINGS
• SCIENTISTS KNEW ABOUT CHROMOSOMES– CHROMOSOMES ARE MADE UP OF• PROTEINS (VERSATILE SUBSTANCES, 20 DIFFERENT
AMINO ACIDS)• NUCLEIC ACIDS (ONLY 5 DIFFERENT NUCLEOTIDES)
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THE MOLECULAR BIOLOGY OF THE GENE
• HERSHEY AND CHASE EXPERIMENT
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THE MOLECULAR BIOLOGY OF THE GENE
• HERSHEY AND CHASE SETUP– USED T2 BACTERIOPHAGES (PHAGE = VIRUS THAT
ATTACKS/EATS BACTERIA)– GREW PHAGES IN RADIOACTIVE ELEMENTS
• SULFUR FOR PROTEIN / PHOSPHOROUS FOR DNA
– TRACING RADIOACTIVITY ALLOWED THEM TO DETERMINE THE GENETIC CARRIER
• RESULTS– RADIOACTIVE PHOSPHOROUS ONLY WAS PRESENT IN
BACTERIAS; CONCLUSIVELY SHOWING THAT DNA (NUCLEIC ACIDS) ARE CONTROLLING TRAITS/GENETICS
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THE MOLECULAR BIOLOGY OF THE GENE
• NOW THE RACE WAS ON TO STUDY DNA/RNA
• DNA / RNA– POLYMERS OF NUCLEOTIDES
• NUCLEOTIDE??• POLYNUCLEOTIDE??
– SUGAR-PHOSPHATE BACKBONE• REPEATING PATTERN
OF SUGAR (PENTOSE) AND PHOSPHATE
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THE MOLECULAR BIOLOGY OF THE GENE
• DNA– DEOXYRIBONUCLEIC
ACID• DEOXYRIBOSE ??• NUCLEIC ??• ACID ??
– 4 TYPES OF BASES IN DNA (2 GROUPS)• PURINES (2 RINGS)
– ADENINE / GUANINE
• PYRIMIDINES (1 RING)
– CYTOSINE / THYMINE
• RNA– RIBONUCLEIC ACID
• RIBOSE ??• NUCLEIC ??• ACID ??
– 4 TYPES OF BASES IN RNA (2 GROUPS)• PURINES (2 RINGS)
– ADENINE / GUANINE
• PYRIMIDINES (1 RING)– CYTOSINE / URACIL
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THE MOLECULAR BIOLOGY OF THE GENE
• STRUCTURE DENOTES ____________– THEREFORE LEARNING THE STRUCTURE OF DNA
BECAME OF EXTREME IMPORTANCE
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THE MOLECULAR BIOLOGY OF THE GENE
• DNA IS A DOUBLE STRANDED HELIX– DOUBLE HELIX TWO STRANDS OF DNA, COILED
AROUND EACH OTHER (*TWISTED LADDER*)– EXPLAINED BY WATSON AND CRICK– USED DATA FROM WILKINS AND FRANKLIN• X-RAY CRYSTALLOGRAPHY
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THE MOLECULAR BIOLOGY OF THE GENE
• WATSON AND CRICK EXPLAINED THE DOUBLE HELIX– USING DATA FROM
ERWIN CHARGAFF (EXPERIMENT THAT SHOWED # OF A’S AND T’S WAS ALWAYS EQUAL, AND # OF C’S AND G’S WAS ALWAYSEQUAL)
– AS WELL AS DATA THAT SHOWED DNA STRANDS WERE ALWAYS THE SAME DISTANCE APART
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THE MOLECULAR BIOLOGY OF THE GENE
• DISCOVERED STRUCTURE SUGGESTED AN EXPLANATION OF DNA REPLICATION
• SPECIFIC BASE PAIRING IS HOW DNA IS COPIED
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THE MOLECULAR BIOLOGY OF THE GENE
• WHILE THE CONCEPT OF REPLICATION IS SIMPLE, ACTUAL PROCESS IS COMPLEX
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THE MOLECULAR BIOLOGY OF THE GENE
• DNA REPLICATION: A CLOSER LOOK– REPLICATION BEGINS AT SPECIFICS POINTS CALLED
ORIGINS OF REPLICATION (O.R.’S)
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THE MOLECULAR BIOLOGY OF THE GENE
• DNA REPLICATION: A CLOSER LOOK– THERE ARE MULTIPLE
O.R.’S ALONG THE DNA STRAND (WHY IS THAT??)
– REPLICATION BUBBLES• WHERE DNA SPLITS AND
REPLICATION OCCURS IN BOTH DIRECTIONS (WHY IS THAT??)
– THE STRANDS OF DNA AREORIENTED IN OPPOSITE DIRECTIONS!!!• HOW WILL THIS AFFECT
REPLICATION? REMEMBER, ENZYMES HAVE SPECIFIC SHAPES!!
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THE MOLECULAR BIOLOGY OF THE GENE
• DNA REPLICATION: A CLOSER LOOK– ENZYMES OF IMPORTANCE• DNA POLYMERASE
– ENZYME THAT ADDS NUCLEOTIDES TO GROW DAUGHTER STRAND
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THE MOLECULAR BIOLOGY OF THE GENE
• DNA REPLICATION: A CLOSER LOOK• DNA LIGASE
– TIES/GLUES PIECES OF DNA TOGETHERINTO A SINGLE CONTINUOUS STRAND
• DNA HELICASE– UNZIPS THE DOUBLE STRANDED HELIX
• TOPOISOMERASE– RELIEVES THE PRESSURE/TENSION PLACED
ON PARTS OF DNA STRAND THAT ARE STILLED COILED
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THE MOLECULAR BIOLOGY OF THE GENE
• DNA REPLICATION: A CLOSER LOOK– ENSURES THAT EXACT COPIES
OF DNA ARE PASSED ALONG TO ALL SOMATIC CELLS
– AMAZINGLY ACCURATE; ONLY ONE MISTAKE EVERY BILLION NUCLEOTIDES PLACED!!
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THE MOLECULAR BIOLOGY OF THE GENE
• THE STRUCTURE OF DNA ALSO OFFERS AN EXPLANATION AS TO HOW IT CAN CONTROL YOUR TRAITS
• THE DNA GENOTYPE (NUCLEOTIDE MAKEUP) IS EXPRESSED AS PROTEINS, WHICH ACTS AS THE MOLECULAR MAKEUP FOR PHENOTYPIC EXPRESSION!!
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THE MOLECULAR BIOLOGY OF THE GENE
• DNA RNA PROTEIN = PHENOTYPE– TRANSCRIPTION• PROCESS BY WHICH DNA IS CONVERTED TO RNA
– TRANSLATION• PROCESS BY WHICH RNA IS CONVERTED TO PROTEIN• PROTEINS CONTROL THE PHENOTYPE (TRAITS) OF AN
ORGANISM
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THE MOLECULAR BIOLOGY OF THE GENE
• BEADLE AND TATUM EXPERIMENT– SHOWED THAT MUTANT
MOLD, DEFICIENT IN ONLY ONE GENE; COULDN’T GROW ON MEDIA THAT IT COULD GROW ON WITH NORMAL GENE
– ONE GENE : ONE POLYPEPTIDE HYPOTHESIS
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSCRIPTION : A CLOSER LOOK– PRODUCES GENETIC MESSAGES
IN THE FORM OF RNA– ONLY OCCURS IN THE NUCLEUS
(WHY??)– SIMILAR TO REPLICATION• 2 STRANDS SPLIT• BUT; ONLY STRAND SERVES AS
TEMPLATE
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSCRIPTION : A CLOSER LOOK– REQUIRES THE FOLLOWING:• RNA POLYMERASE
– ENZYME THAT PLACES AND LINKS NUCLEOTIDES BEING TRANSCRIBED
• PROMOTER– SPECIFIC REGION OF DNA FOR RNA POLYMERASE TO BIND
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSCRIPTION : A CLOSER LOOK– OCCURS IN 3 STAGES• INITIATION• ELONGATION• TERMINATION
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSCRIPTION : A CLOSER LOOK– INITIATION• ????
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSCRIPTION : A CLOSER LOOK– ELONGATION• ????
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSCRIPTION : A CLOSER LOOK– TERMINATION• ?????• TERMINATOR SPECIFIC SEQUENCE ON DNA THAT
SIGNALS THE RNA POLYMERASE TO DETACH
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSCRIPTION : A CLOSER LOOK– EUKARYOTIC RNA IS PROCESSED BEFORE LEAVING
THE NUCLEUS• mRNA = MESSENGER RNA; CARRIES MESSAGE OF DNA
TO RIBOSOME FOR PROTEIN SYNTHESIS• IN EUKARYOTES, THE mRNA MUST BE PROCESSED
(CLEANED UP) BEFORE IT CAN LEAVE
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSCRIPTION : A CLOSER LOOK– EUKARYOTIC mRNA PROCESSING• ONE TYPE OF PROCESSING IS ADDING A “CAP” AND
“TAIL”• WHY DOES THIS HAPPEN??
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSCRIPTION : A CLOSER LOOK– EUKARYOTIC mRNA PROCESSING
• SECOND TYPE OF PROCESSING IS REMOVAL OF NON-CODING REGIONS= RNA SPLICING
• TWO TYPES– EXONS
» ???– INTRONS
» ???
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSCRIPTION : A CLOSER LOOK– EUKARYOTIC mRNA PROCESSING• RNA SPLICING
– REMOVAL OF INTRONS TO PRODUCE FINAL mRNA PRODUCT– SPLICEOSOMES (A.KA. snRNP’S = PRONOUNCED SNURPS)
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSLATION : A CLOSER LOOK– RIBOSOMES BUILD POLYPEPTIDES– RIBOSOMES ARE COMPOSED OF
PROTEINS AND rRNA– RIBOSOMES COORDINATE mRNA,
tRNA, AND AMINO ACIDS TO ALLOW PROTEIN SYNTHESIS
– RIBOSOME STRUCTURE• TWO SUBUNITS (LARGE VS SMALL)• P SITE = PEPTIDYL – tRNA BINDING• A SITE = AMINOACYL – tRNA BINDING
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSLATION : A CLOSER LOOK– tRNA MOLECULES SERVE AS INTERPRETERS DURING
TRANSLATION– AMINO ACIDS ARE
READILY AVAILABLEIN THE CELL FROM DIGESTED FOOD
– tRNA PICKS UP THEAPPROPRIATEAMINO ACID AND BRINGS IT TO THEMATCHING CODON OF THE mRNA
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSLATION : A CLOSER LOOK– CODONS• THREE NITROGENOUS
BASE “WORD” THAT SPECIFIES A PARTICULAR AMINO ACID
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSLATION : A CLOSER LOOK– CODONS– 4 “UNIQUE” CODONS• AUG START CODON, ALSO CODES FOR METHIONINE• 3 STOP CODONS SIGNAL END OF TRANSLATION
– “WOBBLE” HYPOTHESIS• MORE THAN ONE CODON CAN CODE FOR THE SAME
AMINO ACID (OVERLAP)
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSLATION : A CLOSER LOOK
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSLATION : A CLOSER LOOK– TRANSLATION CAN BE DIVIDED INTO THREE
STAGES• INITIATION• ELONGATION• TERMINATION
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSLATION : A CLOSER LOOK– INITIATION
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSLATION : A CLOSER LOOK– INITIATION• 2 STEPS
– mRNA BINDS TO SMALL SUBUNIT OF RIBOSOME; tRNA WITH THE STARD CODON BINDS (MET = AUG)
– LARGE RIBOSOMAL SUBUNIT ATTACHES, CREATING A FUCNTIONAL RIBOSOME; INITIATOR tRNA FITS INTO “P” SITE
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSLATION : A CLOSER LOOK– ELONGATION• ADDS NUCLEOTIDES
TO POLYPEPTIDE CHAIN• 3 STEPS TO
ELONGATION– CODON
RECOGNITION– PEPTIDE BOND
FORMATION– TRANSLOCATION
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THE MOLECULAR BIOLOGY OF THE GENE
• TRANSLATION : A CLOSER LOOK– TERMINATION• ELONGATION CONTINUES UNTIL A “STOP” CODON
ENTERS THE “A” SITE
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THE MOLECULAR BIOLOGY OF THE GENE
• REVIEW
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THE MOLECULAR BIOLOGY OF THE GENE
• MUTATIONS CAN CHANGE THE MEANING OF GENES– MUTATION ANY RANDOM CHANGE IN THE
NUCLEOTIDE SEQUENCE OF DNA– CAN BE CLASSIFIED 3 WAYS• SUBSTITUTIONS• INSERTIONS (ADDITIONS)• DELETIONS
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THE MOLECULAR BIOLOGY OF THE GENE
• SUBSTITUTIONS– A BASE SUBSITUTION
MAY CHANGE AN AMINO ACIDIN A POLYPEPTIDE, CHANGING THE PROTEIN
– “WOBBLE” HYPOTHESIS??
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THE MOLECULAR BIOLOGY OF THE GENE
• INSERTIONS AND DELETIONS– CAN BE MORE SEVERE– THEY CHANGE THE
“READING FRAME”
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THE MOLECULAR BIOLOGY OF THE GENE
• MUTAGENESIS– THE FORMATION OF MUTATIONS– 2 COMMON WAYS
• SPONTANEOUS MUTATION– ERRORS IN DNA REPLICATION/TRANSCRIPTION– UNKNOWN ORIGINS
• MUTAGEN– PHYSICAL (RADIATION) OR CHEMICAL AGENT
• *WHILE MUTATIONS ARE USUALLY HARMFUL; THEY CAN ALSO BE EXTREMELY USEFUL– PROMOTES EVOLUTION– IN THE LAB, PROVIDES A USEFUL TOOL FOR GENETIC RESEEARCH
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THE MOLECULAR BIOLOGY OF THE GENE
• VIRUSES– IN A SENSE, VIRUSES ARE NOTHING MORE THAN
PACKAGED GENES
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THE MOLECULAR BIOLOGY OF THE GENE
• VIRUSES USE HOST CELL MACHINERY TO REPRODUCE– HOW CAN THIS LEAD TO
DISEASE?– WHY ARE VIRUSES DIFFICULT
TO TREAT WITH ANTI-BIOTICS?– HOW DO WE TREAT VIRUSES?
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THE MOLECULAR BIOLOGY OF THE GENE
• VIRUSES– TWO REPRODUCTIVE CYCLES• LYTIC CYCLE
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THE MOLECULAR BIOLOGY OF THE GENE
• VIRUSES– TWO REPRODUCTIVE CYCLES• LYSOGENIC CYCLE **CAN LEAD TO A PROPHAGE
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THE MOLECULAR BIOLOGY OF THE GENE
• EMERGING VIRUSES– AIDS (HIV)– FLU– EBOLA– HANTA
• HOW DO VIRUSES ARISE?– UNKNOWN!!!!– POSSIBLE THEORIES??
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THE MOLECULAR BIOLOGY OF THE GENE
• AIDS– HIV IS A
RETROVIRUS (UNIQUE)
– MAKES DNA FROM AN RNA TEMPLATE
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THE MOLECULAR BIOLOGY OF THE GENE
• AIDS– REQUIRES THE USE
OF REVERSETRANSCRIPTASE
– TURNS HOST CELL INTO A“PROVIRUS”
– WHAT IS THEDIFFERENCEBETWEEN AIDS &HIV??
– WHY IS HIV SO DIFFICULT TO TREAT?
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THE MOLECULAR BIOLOGY OF THE GENE
• THE STRUCTURE OF DNA CAN TELL US HOW IT COPIES ITSELF AND HOW IT CAN GET CONVERTED INTO PROTEIN; BUT HOW COME IF EVERY CELL HAS THE SAME DNA, THEY CAN ALL LOOK DIFFERENT THAN EACH OTHER? YOU JUST HAVE TO STICK AROUND TO FIND OUT MORE!!
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THE MOLECULAR BIOLOGY OF THE GENE