1. You want to clone the gene that codes for a human enzyme you have purified, w
ID: 61919 • Letter: 1
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1. You want to clone the gene that codes for a human enzyme you have purified, which transforms a substrate X into a product Y. Which of the following approaches would be likely to be most successful? use Edman sequencing followed by hybridization of a human genomic library With a non-degenerate probe use Edman sequencing followed by hybridization of a human cDNA library with a degenerate probe Transform a human genomic DNA library into bacteria, and screen for clone that are able to convert X to Y Transform a human genomic DNA library into bacteria, and screen for doesn?t are unable to convert X toy You are given an E. coli mutant strain M that cannot grow without a vitamin Bi. You want to identify the mutated gene. Which of the following approaches would be likely to be most successful? Mix a strain of f - E. coll with M, let conjugation proceed for 1 hour. plate the bacteria on medium containing vitamin Bi, and purify genomic DNA from the colonies. Mix a strain of Hfr E. coll with M, let conjugation proceed for 1 hour, plate the bacteria on medium containing vitamin Bi. and purify genomic DNA from the colonies. Mix a strain of Hfr E. coll with M. let conjugation proceed for 1 hour, plate the bacteria on medium without vitamin Bi, and purify genomic DNA from the colonies. Make a genomic library from wildtype E. coll, transform into M, plate the bacteria on medium without vitamin Bi. and purify library clones from the colonies. Make a genomic library from M DNA, transform into wildtype E. coil, plate the bacteria on medium without vitamin B , and purify library clones from the colonies 3. You need to produce high quantities of pure, elf albumin (NB: elves are supposed to be eukaryotes), and decide to use E. coli as a host (E. coIl does not have any albumin genes). You thus need to create a plasmid suitable for elf albumin expression in E. coli. You do not have a good assay to detect albumin protein. As a starting point to clone the elf albumin gene, you are given the sequence of human albumin (humans and elves are closely related organisms), and a library of your choice. Which of the following DNA libraries would you choose? elf genomic DNA library elf cDNA library E. coll genomic DNA library E coll cDNA library C human genomic DNA library 4. Which of the following methods would you use to identify the albumin gene in your library? cloning by complementation. rescuing E. coil mutants that are defective in albumin synthesis cloning y expression of the library in E. coil and screening clones for presence of elf albumin cloning y hybridization, using the human albumin sequence as a probeExplanation / Answer
1. The correct statement here is "b"
Use edman sequencing followed by hydridization of a human cDNA library with degenerative probe.
3. The correct statement here is "a"
Elf genomic DNA library.
4. The correct statement here is "c"
Cloning by hybridization, using human albumin sequences as probe.
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