An overproducer E. coli strain is an E. coli strain, which carries a recombinant plasmid with an ORF of a protein of interest under the control of a very strong promoter, therefore it produce large quantities of the protein of interest. For example, recombinant human insulin is produced using an overproducer E. coli strain, which carries a plasmid with the ORF of human insulin.

(a) You want to make an overproducer E. coli strain, which will make a protein from bacterium Salmonella, YFPS (your favorite protein from Salmonella). Below is a restriction map of a fragment of Salmonella genomic DNA, which contains ORF of YFPS and a restriction map of a plasmid, which you are using to make the overproducer. Which pair of restriction enzymes would you use?

b.) You want to use the same strategy to produce a human protein in E. coli cells, YFPH (your favorite protein from humans). You have two potential sources to make the insert: a fragment of human genomic DNA, which contains YFPH gene, and a fragment of human cDNA, which contains YFPH ORF. Which one would you use?

c.) What enzyme synthesizes DNA using mRNA as a template during cDNA construction?