THE PROBLEM: You are a scientist in the research and development laboratory of a biotechnology company.You and your colleagues (you may work with up to two additional students on this worksheet) have been tasked with the following biotechnology problem:

THE PROBLEM: You are a scientist in the research and development laboratory of a biotechnology company.You and your colleagues (you may work with up to two additional students on this worksheet) have been tasked with the following biotechnology problem:

• Clone a human gene and mass produce the gene product in a bacterial host (Escherichia coli).

Using your textbook as a resource, complete the following steps to assist you in completing this gene cloning project.

1. You have identified the specific gene that produces the protein product that you wish to mass produce. What is the first step you need to do in order to obtain a version of this human gene that be cloned into a bacterial cell? What special enzyme, obtained from a particular kind of virus, will you need to complete this first step? (Hint: Are there differences in genes between eukaryotes and prokaryotes?)

2. Now that you have your gene available in the correct form, diagram the steps necessary to place the gene’s DNA into a plasmid vector and list all the types of enzymes needed to do this:

3. Once you have created your recombinant plasmid, what process do you use to get the plasmid into your E. coli host cell? (Hint: You have naked DNA in a test tube.)

4. We need to screen our cells to make sure they have the correct plasmid in them. If we cloned using a plasmid containing the lacZ gene at the cloning site, we can use blue/white screening. What color would the cells be that contain a recombinant plasmid?

5. Once you have identified the correct clone containing the gene you wanted, how might you increase the production of the protein product? (Hint: The β-galactosidase gene on your plasmid came from the lactose operon.)