Thinking Like A Biotechnician: Recombinant DNA Lab Analysis

Recombinant DNA

     In this lab, we simulated to process of making Recombinant DNA. Recombinant DNA is the process of inserting the DNA of an organism into the DNA of another organism. First, we identified the gene of interest, which for us was the insulin gene. Then we made a plasmid. A plasmid is a small piece of circular DNA in bacteria. A plasmid is naturally resistant to a certain antibiotic, which for us was tetracycline. Next we used a restriction enzyme. A restriction enzyme cuts DNA wherever it reads a specific sequence. We made sure that the restriction enzyme only cut the plasmid once and the DNA sequence twice. We did this so that there were not to pieces of plasmid and that there was less DNA from the DNA sequence. Once the DNA was cut, we added ligase. Ligase is an enzyme that reattaches the base pairs back together. Finally, we put the bacteria into a petri dish and added the antibiotic tetracycline. We did this to isolate the bacteria that had the insulin gene. The host bacteria produces the gene product, insulin.
     We had a choice of using the antibiotics, tetracycline, kanamycin, and ampicillin. We could only use tetracycline because the DNA segment that had the insulin gene was only resistant to tetracycline. Restriction enzymes cut DNA wherever it reads a specific sequence. We used the restriction enzyme Hpa II because it only cut the plasmid once and the DNA segment twice. If we used an enzyme that cut the plasmid twice, then the plasmid would be in two pieces. This process is important in our everyday life because there are many everyday uses for this technology. Recombinant DNA has allowed scientists to create genetically modified organisms (GMOs), which we commonly eat everyday. This technology has other uses, such as gene therapy, which help people who suffer from genetic diseases. Recombinant DNA technology is still in use today. For example, recently, a new vaccine was made that utilizes a Salmonella bacteria that is genetically modified and has recombinant DNA. There are many uses of Recombinant DNA technology, and we are just scrapping the surface.