ps7q - MIT Department of Biology 7.014 Introductory...

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MIT Department of Biology 7.014 Introductory Biology, Spring 2004 Name:__________________________________________ Section: ______ 7.014 Problem Set 7 Please print out this problem set and record your answers on the printed copy. Problem sets will not be accepted late. Question 1 On your trip to the Amazon you were introduced to a new plant that the indigenous people use as an anti-viral treatment. You took samples home to your lab and you found that this plant makes a protein (the PV protein) that prevents viral replication. Excited by the possible anti-AIDS applications, you construct a genomic DNA library from this plant in the hope of cloning the PV gene. a) What is a genomic DNA library? b) You get a DNA sample from cells, digest it with a restriction enzyme, and clone it into a vector. List 3 features of the vector that are absolutely required for your library construction. c) Circle on the following lists ALL you would need in order to construct the genomic DNA library. Assume you start with intact plant genomic DNA. Enzymes Reagents Restriction enzyme Size separation gel Ligase Okasaki fragments DNA Polymerase ATP, TTP, CTP, GTP RNA Polymerase ddATP, ddTTP, ddCTP, ddGTP Transcriptase Primers Reverse Transcriptase Replication fork 3' to 5' exonuclease E. coli (bacteria) Cloning vector Human cells Virus Briefly describe the function of each item circled. 1
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Name:__________________________________________ Section: ______ Question 1, continued d) Your genomic DNA library is now ready. You have PV protein sequence information and want to make a DNA probe that is 12 nucleotides long. Given the protein sequence below, circle the amino acids that you will use to design your DNA probe. Why did you choose these amino acids? leu-arg-met-gly-ser-ala-val-leu-pro-thr-arg-ser-trp-met-tyr-lys-arg-his-thr-ser-ala-cys-met-pro-phe
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ps7q - MIT Department of Biology 7.014 Introductory...

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