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BIS 140 Mutation Repair

BIS 140 Mutation Repair - (a Transposable genetic elements...

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Unformatted text preview: (a) Transposable genetic elements /\ A B -> C D E —> F G -I-I-I-I-I-I-I-I-I- Ezi‘F III-I- -DI-I- Deletion E (-‘C prod uct ‘I'A B F c -I-III-III- W—l Fig_11-19a Genetics, Second Edition © 2005 W.H. Freeman and Company A B -> C D E " F G (b) -I-I-I-I-I-l-I-I-I- C. F -> E -I-I-I-.‘ D -?-?-_x-%' l A B -> E D C 4- F C _I-I-I-I-I-I-I-l-I- Fig_11-19b Genetics, Second Edition © 2005 W.H. Freeman and Company A B -> C D E -> F G (C) -I-I-I-I-I-I-I-I-I- 0".5 I l .2‘ 'IF (3 I. ‘5 I ‘ c"'D A B F C ------ H—I A B -> C D E " C D E " F G .I-I-I-I-I-I-I-I-I-I-I-I-I- Fig_11-19c Genetics, Second Edition © 2005 W.H. Freeman and Company Table 14-2 Point Mutations at the Molecular Level Type of mutation Result and examples At DNA level Transition Purine replaced by a different purine, or pyrimidine replaced by a different pyrimidine: A~T—>G'C—>G'C—>A'T C'G—>T~A T'A—>C'G Transversion Purine replaced by a pyrimidine, or pyrimidine replaced by a purine: A-T—>C-G A-T—>T-A G-C—>T-A G-C—>C-G T'A—>G'C T-A—>A'T C'G—>A-T C-G—>G'C Indel Addition or deletion of one or more base pairs of DNA (inserted or deleted bases are underlined): AAGACTCCT —> AAGAQCTCCT AAQACTCCT —> AAACTCCT Table 14-2 Point Mutations at the Molecular Level Type of mutation Result and examples At protein level Synonymous mutation Codons specify the same amino acid: AGG —> CGG Arg Arg Missense mutation Codon specifies a different amino acid Conservative missense mutation Codon specifies chemically similar amino acid: AAA —> AGA Lys Arg (basic) (basic) Does not alter protein function in many cases Nonconservative missense mutation Codon specifies chemically dissimilar amino acid: UUU —> UCU Hydrophobic Polar phenylalanine serine Nonsense mutation Codon signals chain termination: CAG —> UAG Gln Amber termination codon Frameshift mutation One base-pair addition (underlined) AAG ACT CCT —>AAG AQC TCC T... One base-pair deletion (underlined) AAQ ACT CCT—>AAA CTC CT... Codon1 Codon2 Codon3 Codon4 I—‘—1|—‘—1I—‘—H—'—1 ACACAGCGTGGT Wild-typegene JENJJ iAcAcAG cGCG Synonymous substitution Base 139 fig} 33') 15y substitutions ACACAGAGTGGT Missense substitution J 3 ‘9 1.9;} 1 Frameshift mutation ACAGCAGCGTG ~- Insertion Regulatory site ACACAGCGTGGTf , i Point mutation ACAGAGCGTGGT innoncoding |——' sequence Regulatory protein cannot bind / ."H Cytosine \N | \ \ H Guanine Adenine Thymine O O..H\ N \ Rareenol form | Adenine ofthymine(T*) H Guanine Rare imino form of cytosine (C*) \ CI-I3 0.. 'H\ Rare Imino form of adenine (A*) H Cytoosine H Rare enol form of guanine (G*) Common keto Adenine Rare Guanine form of 5-BU ionized form of 5-BU (a) (b) 2-AP Thymine Protonated Cytosine 2-AP (a) (b) Guanine O-6-Ethylguanine Thymine if i “rs-W \H Thymine O-4-Ethylthymine Guanine H\N,CH2CH2CI I /CH2 H CPI/CH2 2 \N/ OCH3 Proflavin Acridine orange lCR-191 Nitrogenous bases lntercalated molecule . Stalled . Base I Error-prone DNA replication machine damage polymerases bind stalled replication machine. ' J Location marker POI 1] P°l 'v to show movement I Binding promotes conformational change. Replication begins. pol 1] adds erroneous bases. Pol 1] disassociates. pol I. continues incorporating erroneous bases on strand opposite damaged base. (a) Direct reversal W (b) Base excision and replacement MW (c) Segment removal and replacement MW 0’ P\ Cyclobutyl ring f”: I0 0\ HZC [P :00 lo 0 H ,CH2 3' 0 ‘P ,0 o / ‘ I 0 ° 0; P (a) Cyclobutane pyrimidine dimer 5' ‘o (b) 6-4 Photoprocluct Thymine Thymine DNA backbone Photolyase + white Photodimer light UV light W AP endonuclease makes cut WEWEEE Excision exonuclease removes stretch of DNA WE iiagi Polymerase synthesizes new DNA WEE? Ligase seals nick mm 5' E 3' 3' 5' uvrABC excinuclease removes a 12-nucleotide fragment of DNA W DNA polymerase I synthesizes new DNA W Ligase joins DNA segments W Repairosome TFIIH subunit unwinds DNA damaged strand 1 Excision of 5' incision DNA synthesis and ligation Velvet surface (sterilized) Handle ( ? —> Pressed on master Then pressed on replica plate containing plate that distinguishes wild grown colonies and mutant genotypes Master plate containing 107 colonies of Ton 5 E. coli (T1-sensitive) . O . O . O O O . Plate-1 Plate '2 Plate 3 Series of replica plates containing high concentrations of T1 phage and four Ton ' colonies ...
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