MCB121 W09 MT1practice exam

MCB121 W09 MT1practice exam - Name _ MCB121 W08 MTI A...

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Name ___________________________ MCB121 W08 MTI A February 5, 2008 1 ID #_______KEY_________________________ FIRST MIDTERM February 5, 2008 Question Points Score 1 24 2 20 3 12 4 20 5 20 6 12 7 6 8 6 9 4 10 6 11 20 Total 150 This examination is closed book. There are 9 pages to the exam. Please count them before you start to make sure all are present . Please write your name on each page of the exam . Answer each question in the space provided. If additional space is required use the back of the page and indicate clearly that you continued your answer on the back. Do not attach additional pages. GOOD LUCK! This is a practice exam from last year. Questions covering topics I have not discussed yet this year are shaded. -SB Student authorization: I authorize the instructor to return the graded exam to me in the main office or to distribute it in the bin for me to pick up.
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Name ___________________________ MCB121 W08 MTI A February 5, 2008 2 Signature _____________________________
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Name ___________________________ MCB121 W08 MTI A February 5, 2008 3 1. (24 points) The plasmid map below is of an expression vector (pNE1) for producing foreign proteins in E. coli . The “Tac Promoter” is a strong promoter for transcription of the adjacent region. The sequence adjacent to the Tac promoter is shown below the map. “BamHI”, "NcoI" and "EcoRI" indicate recognition sequences for three different restriction endonucleases (these are capitalized and underlined in the sequence and are the only BamHI, NcoI and EcoRI sites in the vector). RBS is a ribosome binding site, and the ATG start codon for expression of proteins by this vector is bold and underlined in the sequence. Construct: pNE1 (2259 bp) BamHI RBS NcoI EcoRI Tac Promoter BamHI RBS NcoI EcoRI_ …aaGGATCC atattatacgGGAGGcctcCC ATG G ggctccctagctcagtGAATTC cc… You want to insert the coding region from the sequence below, encoding a protein called FKBP, into this vector so that the intact protein will be produced. The ATG start codon and TGA stop codon of the protein coding region are capitalized and underlined. ccgcccgccc gctcagcgtc cgccgccgaa ATG ggagtgc aggtggaaac catctcccca ggagacgggc gcaccttccc caagcgcggc cagacctgcg tggtgcacta caccgggatg cttgaagatg gaaagaaatt tgattcctcc cgggacagaa acaagccctt taagtttatg ctaggcaagc aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca aactgactat atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac cacatgccac tctcgtcttc gatgtggagc ttctaaaact ggaa TGA cag gaatggcctc ctcccttag
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Name ___________________________ MCB121 W08 MTI A February 5, 2008 4 A. (4 points) To insert the coding sequence you have decided to add restriction enzyme recognition sequences at the two ends of the coding region using the polymerase chain reaction (PCR). What restriction enzyme recognition sequences would you add near the start and stop codons of the coding region? Near start – Near stop – B. (12) Design primers that have at least 10 bases of overlap with the gene sequence that you could use for PCR to produce a fragment that could be appropriately inserted into the pNE1 expression vector. Write your primer sequences below going from 5’ to 3’ of each primer sequence .
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This note was uploaded on 04/07/2010 for the course MCB 102,103,12 taught by Professor Segel during the Spring '10 term at UC Davis.

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MCB121 W09 MT1practice exam - Name _ MCB121 W08 MTI A...

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