ProblemSet8

ProblemSet8 - Biology 202 From Griffiths: Problems Problem...

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Biology 202 Problem Set 8 4/16/10 From Griffiths: Problems Chapter 20: 1, 4, 5, 6, 10, 18, 19 Answers to the following questions are due April 23 th at 4:00 PM. 1. Some restriction enzymes have degenerate recognition sequences. For example, Sty I cuts at the sequence 5'-CCWWGG-3', where W = A or T (W stands for weak, because A–T has only two H-bonds). a) Which of the following sequences will be cut by Sty I? 5’–CCAAGG–3’ 5’–CCTAGG–3’ 5’–CCGAGG–3’ 5’–GGTTCC–3’ 3’–GGTTCC–5’ 3’–GGATCC–5’ 3’–GGCTCC–5’ 3’–CCAAGG–5’ b) Sty I cuts between the two C’s on each strand. The enzyme Nco I cuts at the sequence 5'-CCATGG-3', between the two C’s on each strand. What fraction of sites cut with Sty I will give ends that can be ligated to DNA cut with Nco I? Will the product contain a site for Sty I? For Nco I? c) The enzyme Ocn I cuts the sequence 3'-CCATGG-5' (note that this is written 3’ to 5’) between the two C’s on each strand. Can DNA cut with Ocn I be ligated to DNA cut with Nco I? Explain. d) The enzyme Nut I cuts at the sequence 5'-CCATGG-3', between the A and T. Can DNA cut with Nut I be ligated to DNA cut with Nco I? Explain. 2. As a scientist, you have decided to make some recombinant DNA in the hopes that you will find a cure for a rare genetic disease. Your colleagues--who are familiar with recombinant DNA techniques--encourage you to use cDNA as your donor DNA. a) What is cDNA and how does it differ from a genomic DNA molecule?
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b) Discuss the advantages and disadvantages of using cDNA as your donor. 3. Human insulin can be made purer, and at a lower cost, using recombinant DNA technology. What sort of donor DNA is used in the creation of bacteria that express human insulin. Explain why. 4. The restriction enzyme BglII (from Bacillus globigii) cuts in the following place (*) 5'-A*-G-A-T-C-T-3' a) Draw the complementary strand and indicate where BglII will cleave the DNA. b) How would you characterize the fragments? c) The restriction enzyme Hae III cleaves in the following place (#). Compare the DNA fragments that exist after the restriction enzyme has made its cut to those in part (a). 5'-G-G #-C-C-3' 5. You have written out a protocol for cloning a gene, and are assembling the supplies you need to have in the lab. Look over the list below. Do you have all the necessary supplied listed? If not, add the supplies you would need and what they would be used for: a) Donor DNA amplified with PCR primers that added BamHI and HindIII sites at either end. b) Vector DNA: contains the ampicillan resistance gene, a multicloning site with
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This note was uploaded on 04/20/2010 for the course BIOL 202 taught by Professor Kieber-hogan during the Fall '08 term at UNC.

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ProblemSet8 - Biology 202 From Griffiths: Problems Problem...

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