202hw10KEY - Biology 202 Homework 10 KEY Due Friday Dec 5...

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Biology 202 Homework 10 KEY Due: Friday, Dec. 5, by 5:00 p.m. 1. You have received a cloned DNA segment (in a plasmid) and determined that the insert is 1300 base pairs (bp) long. Using the restriction enzymes Dpn II and Hae III, followed by gel electrophoresis, you determine the sizes of fragments produced by these enzymes separately and together as shown in the table below: ENZYME RESTRICTION FRAGMENT SIZES Dpn II 350 bp, 950 bp Hae III 200 bp, 1100 bp Dpn II and Hae III 150 bp, 200 bp, 950 bp Construct a linear restriction map (a map that indicates the location of restriction sites) from these data showing the positions of restriction sites relative to one another and the distance between them in base pairs. (2 pts) HaeIII DpnII |_____200 ___|__150 ___|_____________________________950 __________________________________________| 2. Most E. coli have a naturally low mutation rate where the probability of a particular base substitution is on the order of 10 -10 per base pair in every generation. However, some naturally occurring pathogenic strains have a mutation rate that is orders of magnitude higher than that of the wild type. You are interested to know whether this large difference in mutation rate between strains could be due to mutations in a gene for a key mismatch repair protein, MutS. For this purpose you would like to sequence the mutS gene from strains with high mutation rate and compare those sequences with mutS from strains with low mutation rates. a. Before you can sequence this gene, you will need large quantities of DNA for this gene and you plan to use the polymerase chain reaction (PCR) for that. You know the sequence of mutS from a laboratory strain of E. coli that has a low mutation rate, and the first 50 and the last 50 nucleotides of the non-template (coding) strand of the gene is shown
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This note was uploaded on 01/29/2009 for the course BIOL 202 taught by Professor Staff during the Spring '08 term at New Mexico.

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202hw10KEY - Biology 202 Homework 10 KEY Due Friday Dec 5...

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