A) Transformation is a genetic process which means change caused by genes, and involves the insertion of foreign DNA into an organism to change its trait. Transformation is carried out by restriction enzymes to cut plasmid DNA and insert new code in its place, and heat shock (rapidly heating and cooling cells with calcium chloride) which will move the plasmid (DNA which contains one or more genes which would be beneficial to a bacteria's survival) into the cell. It also is a key instrument of biotechnology, and is used in numerous ways, such as, bioremediation (the genetic manipulation of bacteria to digest oil from spills), and medicine (use of gene therapy to correct defective genes). Also, another use of Transformation is to make bacteria glow by inserting the Green Fluorescent Protein (from Seajellies) into bacterial cells.
B) The purpose of this experiment is to insert the GFP gene into a plasmid, and move the plasmid into bacterial cells. This will cause the cells to produce Green Fluorescent Protein, and they will glow brilliantly. We hope to have a flourishing fluorescent colony by the end of this experiment.
C) In this lab, we will have four petri dishes: +pGLO and LB/amp, -pGLO and LB/amp, -pGLO, and LB, and the +pGLO and LB/amp/ara. We will use Transformation, which is carried out by restriction enzymes to cut plasmid DNA and insert new code (GFP gene) in its place, and heat shock (rapidly heating and cooling cells with calcium chloride) which will move the plasmid (DNA which contains one or more genes which would be beneficial to a bacteria's survival) into the cell. The controls of this experiment are the -pGLO, and LB petri dishand the -pGLO and LB/amp. The variables will be +pGLO and LB/amp,and the +pGLO and LB/amp/ara. I predict that the +pGLO and LB/amp will be the only bacterial colony to exhibit fluorescent characteristics. This is because it contains the ampicillin resistance gene, and the plasmid containing the GFP gene.
