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THE MOLECULAR BIOLOGY OF THE GENE

THE MOLECULAR BIOLOGY OF THE GENE. 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

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THE MOLECULAR BIOLOGY OF THE GENE

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  1. THE MOLECULAR BIOLOGY OF THE GENE

  2. 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?)

  3. THE MOLECULAR BIOLOGY OF THE GENE • GRIFFITH’S EXPERIMENT

  4. 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

  5. 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)

  6. THE MOLECULAR BIOLOGY OF THE GENE • HERSHEY AND CHASE EXPERIMENT

  7. 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

  8. 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

  9. 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

  10. THE MOLECULAR BIOLOGY OF THE GENE • STRUCTURE DENOTES ____________ • THEREFORE LEARNING THE STRUCTURE OF DNA BECAME OF EXTREME IMPORTANCE

  11. 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

  12. 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

  13. THE MOLECULAR BIOLOGY OF THE GENE • DISCOVERED STRUCTURE SUGGESTED AN EXPLANATION OF DNA REPLICATION • SPECIFIC BASE PAIRING IS HOW DNA IS COPIED

  14. THE MOLECULAR BIOLOGY OF THE GENE • WHILE THE CONCEPT OF REPLICATION IS SIMPLE, ACTUAL PROCESS IS COMPLEX

  15. THE MOLECULAR BIOLOGY OF THE GENE • DNA REPLICATION: A CLOSER LOOK • REPLICATION BEGINS AT SPECIFICS POINTS CALLED ORIGINS OF REPLICATION (O.R.’S)

  16. 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 ANDREPLICATION 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!!

  17. THE MOLECULAR BIOLOGY OF THE GENE • DNA REPLICATION: A CLOSER LOOK • ENZYMES OF IMPORTANCE • DNA POLYMERASE • ENZYME THAT ADDS NUCLEOTIDES TO GROW DAUGHTER STRAND

  18. 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 PLACEDON PARTS OF DNA STRAND THAT ARE STILLED COILED

  19. 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!!

  20. 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!!

  21. 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

  22. 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

  23. 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 ASTEMPLATE

  24. 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

  25. THE MOLECULAR BIOLOGY OF THE GENE • TRANSCRIPTION : A CLOSER LOOK • OCCURS IN 3 STAGES • INITIATION • ELONGATION • TERMINATION

  26. THE MOLECULAR BIOLOGY OF THE GENE • TRANSCRIPTION : A CLOSER LOOK • INITIATION • ????

  27. THE MOLECULAR BIOLOGY OF THE GENE • TRANSCRIPTION : A CLOSER LOOK • ELONGATION • ????

  28. THE MOLECULAR BIOLOGY OF THE GENE • TRANSCRIPTION : A CLOSER LOOK • TERMINATION • ????? • TERMINATOR  SPECIFIC SEQUENCE ON DNA THAT SIGNALS THE RNA POLYMERASE TO DETACH

  29. 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

  30. 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??

  31. 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 • ???

  32. 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)

  33. THE MOLECULAR BIOLOGY OF THE GENE • TRANSLATION : A CLOSER LOOK • RIBOSOMES BUILD POLYPEPTIDES • RIBOSOMES ARE COMPOSED OFPROTEINS 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

  34. THE MOLECULAR BIOLOGY OF THE GENE • TRANSLATION : A CLOSER LOOK • tRNA MOLECULES SERVE AS INTERPRETERS DURING TRANSLATION • AMINO ACIDS AREREADILY AVAILABLEIN THE CELL FROM DIGESTED FOOD • tRNA PICKS UP THEAPPROPRIATEAMINO ACID AND BRINGS IT TO THEMATCHING CODONOF THE mRNA

  35. THE MOLECULAR BIOLOGY OF THE GENE • TRANSLATION : A CLOSER LOOK • CODONS • THREE NITROGENOUS BASE “WORD” THAT SPECIFIES A PARTICULAR AMINO ACID

  36. 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)

  37. THE MOLECULAR BIOLOGY OF THE GENE • TRANSLATION : A CLOSER LOOK

  38. THE MOLECULAR BIOLOGY OF THE GENE • TRANSLATION : A CLOSER LOOK • TRANSLATION CAN BE DIVIDED INTO THREE STAGES • INITIATION • ELONGATION • TERMINATION

  39. THE MOLECULAR BIOLOGY OF THE GENE • TRANSLATION : A CLOSER LOOK • INITIATION

  40. 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

  41. THE MOLECULAR BIOLOGY OF THE GENE • TRANSLATION : A CLOSER LOOK • ELONGATION • ADDS NUCLEOTIDESTO POLYPEPTIDE CHAIN • 3 STEPS TOELONGATION • CODONRECOGNITION • PEPTIDE BONDFORMATION • TRANSLOCATION

  42. THE MOLECULAR BIOLOGY OF THE GENE • TRANSLATION : A CLOSER LOOK • TERMINATION • ELONGATION CONTINUES UNTIL A “STOP” CODON ENTERS THE “A” SITE

  43. THE MOLECULAR BIOLOGY OF THE GENE • REVIEW

  44. 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

  45. THE MOLECULAR BIOLOGY OF THE GENE • SUBSTITUTIONS • A BASE SUBSITUTION MAYCHANGE AN AMINO ACIDIN A POLYPEPTIDE, CHANGING THE PROTEIN • “WOBBLE” HYPOTHESIS??

  46. THE MOLECULAR BIOLOGY OF THE GENE • INSERTIONS AND DELETIONS • CAN BE MORE SEVERE • THEY CHANGE THE “READING FRAME”

  47. 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

  48. THE MOLECULAR BIOLOGY OF THE GENE • VIRUSES • IN A SENSE, VIRUSES ARE NOTHING MORE THAN PACKAGED GENES

  49. THE MOLECULAR BIOLOGY OF THE GENE • VIRUSES USE HOST CELL MACHINERY TO REPRODUCE • HOW CAN THIS LEAD TODISEASE? • WHY ARE VIRUSES DIFFICULTTO TREAT WITH ANTI-BIOTICS? • HOW DO WE TREAT VIRUSES?

  50. THE MOLECULAR BIOLOGY OF THE GENE • VIRUSES • TWO REPRODUCTIVE CYCLES • LYTIC CYCLE

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