BIMM100 Problem Set 2_JL_TAs

4 product 5 d you repeat the reaction above but now

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Unformatted text preview: de the primer sequence in the product). (4) Product: 5’- d) You repeat the reaction above, but now include a small amount of dideoxythymidine triphosphate (ddTTP), as in a Sanger sequencing reaction. Write, in the 5’-to-3’ direction, all of the products you would get after DNA synthesis from the primer (You do not need to write the template). (4) Products (5’-to-3’): Question 9. 45% of our genome is mobile elements, which come in two flavors: DNA transposons and Retrotransposons. Below is shown the steps in transposition of IS10, with DNA intermediates/ products numbered [1-4]. a) If IS10 transposition was trying to take place in the absence of the activities listed below, for each indicate which DNA intermediate or product would accumulate [1, 2, 3, or 4]: (4) 1 - Transposase (encoded by IS10): - Reverse Transcriptase: - DNA polymerase: - Restriction enzyme: 2 b) When transposition occurs into the promoter, 5’ UTR, coding region or 3’UTR of an important protein-coding gene, it can create a problem for the cell. For each of these possible occurrences, indicate whether the transposition is likely [yes/no] to alter the amino acid sequence of the protein produced by the affected gene. (4) Insertion into Promoter: 5’UTR: Coding region: 3’UTR: 3 4 c) Below is shown a schematic of IS10 having inserted itself into the 5’UTR of gene X. You wish to develop a PCR-based method for genetic screening for this insertion. On the schematic, indicate where you would place the PCR primers, clearly indicating 5’-to-3’ directions of the primers and whether the primers should be inside or outside of the IS10 sequence. (4) Gene X Promoter 5’UTR coding region 3’UTR IS10 d) Some pseudogenes have arisen when retrotransposons mistakenly retrotranspose a normal gene instead of the retrotransposon. In two sentences or less, explain why this is associated with loss of the gene’s introns. (4) Question 10. You wish to express a human protein in E. coli. You...
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This note was uploaded on 11/04/2013 for the course BIMM 100 taught by Professor Pasquinelli during the Fall '06 term at UCSD.

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