Exam 1 2008 BLANK - 1 Biochemistry 441 Hour Exam #2 March...

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1 Biochemistry 441 Hour Exam #2 March 3, 2008 Name (First, last) _______________________________________ Student number _______________________________________ Page 1 of 10 Please count your pages to be sure you have the correct number. Question Value Score 1 6 ______________ 2 5 ______________ 3A,B 7 ______________ 3C 6 ______________ 3D 8 ______________ 3E 8 ______________ 4A,B 10 ______________ 4C,D 10 ______________ 5A,B 10 ______________ 6A,B 10 ______________ 7 7 ______________ 8 7 ______________ 9 6 ______________ Total 100 ______________
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2 1. ( 6 points). Which technique listed below (matching terms a-f) would be the most appropriate to use for the purposes stated in lines 1-6? 1. Detect a DNA binding protein in cell extracts __ c ____ 2. Determine the 5’ end of a mRNA ___ d ____ 3. Identify all of the genes transcribed in brain tissue ___ f ____ 4. Accurately measure the amount of mRNA produced by a specific gene __ a __ 5. Identify the binding site for a purified transcription factor ___ e ___ 6. Demonstrate that a specific transcription factor binds to the promoter of a particular gene in vivo ___ b ___ Matching terms (use each term only once): a. quantitative “real-time” reverse transcriptase PCR b. chromatin immunoprecipitation c. electrophoretic mobility shift assay (gel shift assay) d. S1 nuclease assay e. DNase footprinting f. DNA microarray 2. (5 points) Match the most appropriate term listed in a-e with phrases 1-5. 1. located at the 5’ end of most eukaryotic mRNAs. ___ d ___ 2. includes a 2’, 5’ phosphodiester bond. ___ b ___ 3. allows the rapid purification of eukaryotic mRNAs. ___ c ___ 4. increases the types of proteins that a cell can produce. ___ a ___ 5. complementary to the 5’ splice site. ___ e ___ Matching terms (use each term only once): a. alternative splicing b. lariat structure c. poly(A) tail d. 7-methyl guanosine e. U1 snRNA
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3 3. Archaebacteria are single-celled microbes thought to constitute a kingdom of life distinct from both eukaryotes and true bacteria (eubacteria). The following questions relate to the transcription apparatus in the archaebacterium Thermus aquaticus (the source of taq polymerase). A. (3 points) You want to purify RNA polymerase from a mixture of proteins extracted from T. aquaticus cells. In order to detect the activity of RNA polymerase, what components would you have to add to the reaction mixture in addition to the proteins? Template DNA, all four NTPs, and Mg ++ B. (4 points) When an RNA polymerizing activity from T. aquaticus was purified and analyzed by SDS polyacrylamide gel electrophoresis, the results shown in Figure 1 were obtained. If all these polypeptides are part of a T. aquaticus holoenzyme, is the enzyme more similar to the RNA polymerase from eubacteria or yeast? Briefly explain your answer. It is more similar to eukaryotic RNA polymerases because it has 11 subunits.
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Exam 1 2008 BLANK - 1 Biochemistry 441 Hour Exam #2 March...

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