section18 - MIT Department of Biology 7.014 Introductory...

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7.014 Introductory Biology, Spring 2005 Recitation Section 18 April 13-14, 2005 Recombinant DNA—Libraries and Forensics A. Library construction Recall that earlier in the semester we considered the C ys4 yeast gene that encodes a CBS protein. Recall also that yeast mutant in Cys4 are cysteine auxotrophs, while humans mutant in a homologous gene have a host of medical problems from mental retardation to hear disease and ocular degeneration. Today we will talk about finding the yeast gene responsible. In Section 22 we will turn to the question of using a yeast system to diagnose this particular human disease. Suppose we wanted to find the particular gene responsible for the cysteine auxotrophy phenotype. We decide to use cloning by complementation, and we use one of our cysteine auxotroph cys4 - mutant strains as a host strain. 1. What is a library? 2. What is a library used for? 3. What is a common principle between complementation by mating that we did in Genetics and identification of a gene using genomic library? We need a vector and a source of genomic DNA to construct a library. 4. What must the genotype at the Cys4 locus be of the strain of yeast that we use as the source of genomic DNA? What phenotype must it have? Why? 5. What features should the vector we are using have?
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This note was uploaded on 01/19/2012 for the course BIOL 7.014 taught by Professor Walker during the Spring '05 term at MIT.

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section18 - MIT Department of Biology 7.014 Introductory...

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