MCB121 PMT2 Winter 2011

MCB121 PMT2 Winter 2011 - Name Last First MCB121 Page 1 of...

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Name MCB121, Page 1 of 10 Last, First Gasser, Burgess MCB121 Second Midterm, Feb 18, 2010 Instructions: There are nine pages in this exam including the cover sheet, please count them before you start to make sure all are present. Write your name on each page of the exam. Write your answers in the space provided below each question. If you need more space use the back of the page and indicate clearly that you have continued your answer on the back. Do not use additional paper. Amino acid Abbreviations Amino acid Abbreviations Alanine Ala A Leucine Leu L Arginine Arg R Lysine Lys K Asparagine Asn N Methionine Met M Aspartic acid Asp D Phenylalanine Phe F Cysteine Cys C Proline Pro P Glutamine Gln Q Serine Ser S Glutamic Acid Glu E Threonine Thr T Glycine Gly G Tryptophan Trp W Histidine His H Tyrosine Tyr Y Isoleucine Ile I Valine Val V 1. (20 points) For each of the following, write what kind of organism (yeast, mammals, or plants – more than one if more than one applies) it has utility for in eukaryotic transformation and briefly describe how it is useful. (5 points each) a) Gene replacement b) Embryonic stem cells c) Agrobacerium tumefaciens d) Retroviral vector
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Name MCB121, Page 2 of 9 Last, First Gasser, Burgess 2. (12 points) Mark the single most correct answer to each of the following. a) (4) Which of the following is NOT a feature of a human cDNA library? The absence of introns allows for easy identification of critical promoter sequences in the clones. Reverse transcriptase is necessary for library construction. Libraries made from different organs of the same individual would contain significantly different populations of clones. It can include clones representing the population of mRNA in a cell or tissue sample. When inserted into an appropriate expression vector it could be effectively screened in bacteria using an activity assay for a particular human enzyme. b) (4) A transient expression analysis of a cloned gene in cultured mouse cells: Works best if telomeres are included in the transforming DNA. Requires a promoter that functions in mouse cells to be effective. Can only be done for mouse cDNA clones because genomic copies of mouse genes include introns. Depends on a positive selectable marker to ensure that the gene is present in the analyzed cells. Depends on a negative selectable marker to eliminate non-transformed cells. c) (4) Which of the following would NOT be an important feature to include in a yeast artificial chromosome (YAC) cloning vector? A centromere sequence. A selectable marker for yeast cells. A telomere region. An intron. A promoter region active in yeast.
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Name MCB121, Page 3 of 9 Last, First Gasser, Burgess 3. (12 points) The map below shows the region of an expression vector that includes sequences for expression of inserted coding regions. Also included in the vector are a 6- His region encoding six histidine residues and an adjacent region encoding the “Myc” peptide, both of which can be used for fusions with the protein encoded by an inserted
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MCB121 PMT2 Winter 2011 - Name Last First MCB121 Page 1 of...

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