Recombinant DNA Lab Analysis

Recombinant DNA Lab Analysis

In this lab, we made "recombinant DNA" models out of paper. Recombinant DNA is created through a process called transformation. First, a "gene of interest" is located, finding the location and sequence of the gene and its surrounding sequences. In our case, this gene was called the insulin gene. During transformation,  Enzymes called Restriction Enzymes cut the DNA when it reaches a specific sequence of bases. Different enzymes cut at different sequences. We used an enzyme called Eco RI, because it cut our plasmid once and our DNA twice, close to the gene of interest. This is important because it allows us to splice in the insulin gene to the plasmid. If we used an enzyme which cut the plasmid in two places, then the plasmid would be split into two pieces and we wouldn't be able to put in the gene. When it cuts it, it leaves a "sticky end," which helps the DNA bond with other DNA or plasmids. Plasmids are rings of DNA most commonly found in prokaryotes. Usually they have genes in them which give antibiotic resistance. Our plasmid contained a gene for resistance to both Tetracycline (used to treat acne and skin infections) and Kanamycin (treats serious bacterial infections). Ligase, an enzyme which puts base pairs back together, bonds the cut plasmid and DNA. Then the recombinant plasmid and bacteria are mixed, and the non-resistant bacteria are weeded out by adding whichever antibiotic the recombinant plasmids are resistant to.  For example, we would add in either Tetracycline or Kanamycin, because if the bacteria have taken in our plasmid, which has resistance to both, than it will survive. If it doesn't, it won't. We wouldn't use an enzyme such as ampicillin, because our plasmids are not resistant to it.  After the mixing, the gene is extracted and purified. This process is important in our everyday life because it allows us to mass produce necessary items such as insulin and more. This process could be used for herbicide or pesticide resistant crops or to clone organisms as well.

This picture is of our "recombinant plasmid" and the different restriction enzymes.