1. To express a mammalian gene in bacteria, should the promoter be of bacterial origin or mammalian origin and why?
2. List 3 features that are essential for a plasmid to function as a cloning vector?
3. Given a 1kb gene cloned into a plasmid vector, what are two ways by which you can generate large amounts of this gene?
4. If you want to identify genes from human liver that are highly expressed only under conditions of high stress, what kind of DNA library will be the best one to use and why? How will you construct this library?
5. Why do Type II Restriction Endonucleases not degrade bacterial chromosomal DNA?
6. If a double-stranded circular DNA molecule has 4 recognition sites for HindIII, how many fragments will be generated if you digest that molecule with HindIII?How many fragments will result if you linearize the above ds DNA molecule and then digest with HindIII? describe!
7. If you are tasked with screening a cDNA expression library for the production of a specific protein, what kind of library screening method will you use and why?
1. A biotech company has obtained tissue from normal mice and from mice that has especially engineered, highly active muscle cells. You are hired by the company to identify the genes (mRNA) that are differentially expressed in the normal versus the engineered mice. describe in one page how you will design an experiment to complete your assignment. Your response should specifically address the problem.
2. Suppose you have successfully isolated the putative gene that confers resistance to viral flu, but now you need to show that the gene does actually protect you from getting the flu. Briefly, describe two different ways by which you can do the functional characterization of the putative viral flu gene that you have isolated, in a mouse model?
3. Let's say you have two bacterial samples in tubes 1 and 2, one of which has been transformed with ONLY a vector and the other with a RECOMBINANT construct. Using ONLY two techniques-PCR and gel electrophoresis (use both), how will you determine which tube carries a recombinant and which tube the vector alone? Describe where the primers will anneal (not specific sequence but general location), what you expect to observe on the gel and how you will analyze and draw conclusions from the results? A schematic may be helpful