1.     What are plasmids? Where are they found? Why are they important to the practice of genetic engineering? 

2.     Do plasmids have an importance beyond the practice of genetic engineering? Explain.

3.     What are restriction enzymes?

4.     You might wonder why we might have and origin of replication indicated on the plasmid. What is the origin of replication and why is it important to the genetic engineering process?

5.     You might wonder why there are antibiotic resistance genes in the plasmid [genes that codes for resistance to specific antibiotics). Hint: “The antibiotic resistance genes will be used for screening purposes.” What could this mean? Explain.

6.     Why would you want your restriction enzyme to cut as close as possible to the insulin gene without cutting into it?

7.     [IMPORTANT: YOUR RESPONSE TO THIS QUESTION #7 WILL BE SUBMITTED AS A SEPARATE DOCUMENT] The Human Insulin Gene is a sequence of DNA that ultimately codes for amino acids of specific identities in a specific order.  Since DNA is a double-stranded molecule, there are two complementary sequences present: the template strand and the coding strand. From the Human Insulin Gene sequence in your RECOMBINANT PLASMID, you should be able to generate a list of the appropriate amino acids in the correct order. All you will need is a codon chart; a chart that cross-indexes codons with amino acids (OpenStax Concepts of Biology, Figure 9.20, p.220). You will submit this response (#7) on a separate piece of paper with “#7 HUMAN INSULIN: AMINO ACID SEQUENCE” centered at the top of the page. The amino acids should be listed in a vertical column (or columns, if needed). Be sure to list the amino acids by their standard three-letter abbreviation. For example, your first amino acid will be Met (methionine).