Biol 385: Biotech. and Gen. Eng. Name ___________________________
Exam #2 - 100pts 06 April 2012 G # _____________________
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If necessary, use the back of the exam pages for the rest of your answers. Do not use other sheets of paper. Please write legibly; if I cannot read your answer, I will count it wrong. * BY TAKING THIS EXAM, YOU HAVE AGREED TO ABIDE BY THE SPIRIT AND THE LETTER OF THE HONOR CODE OF GEORGE MASON UNIVERSITY. INITIAL HERE
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1. (6pts) Expression of eukaryotic proteins in prokaryotic cells occasionally results in instability or
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Draw a schematic of the vector and include all of the genetic loci that will permit this two-gene vector to function as a shuttle vector between human cells and E.coli cells. (For your convenience, I have put the A and B polypeptide coding regions on the two-gene vector schematic.)
See Figure 7.20 on page 184, Chapter 07 of the text.
A two gene vector will have a two promoters and two polyA sites to permit expression of both coding regions, which will be on two mRNAs. As a shuttle vector it will also have an E.coli ori and selectable marker, and a eukaryotic selectable marker and either a eukaryotic ori or sequences to permit homologous recombination into the chromosome.
7. (3pts) A technician from the BIOtique, Ltd. Protein Engineering Group comes to you for advice regarding her protein of interest. She needs to improve the enzyme in order for Biotique, Ltd. to compete effectively in a new commercial market area. What are three properties of her enzyme that you could suggest that might be useful to change by directed mutagenesis in order to improve her enzyme?
1) kinetic property - Vmax kinetic property - Km Thermal stability pH optimum
2) altered allosteric regulation increased resistance to degradation removal of co-factor requirement increased reactivity in non-aqueous solvent
3) altered specificity of substrate binding
8. (6pts) BIOtique, Ltd.’s Molecular Genetic Diagnostics
The highlighted red answers are the ones that are correct. The simplest way of navigating through this
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• Use these data to construct a map of these three genes in two steps.
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