E.coli Transformation
Brooke Rowlett
November 26, 2013
BioL 230 Lab

Introduction In this experiment we attempted to transform e.coli using a green fluorescent protein plasmid. This green fluorescent protein is naturally found within the bioluminescent jellyfish. The protein can be expressed in both prokaryotic and eukaryotic cells. Normally whenever bacterial cells contain this protein are exposed to long wave UV radiation, they emit a green light. Bacterial cells acquire the glowing ability through the transformation of the bacteria by a plasmid containing the GFP gene. Transformation is the process in which bacteria take up exogenous DNA to acquire to traits (Witucki). The GFP would be our evidence to see whether or not the transformation was successful. Within this experiment E.coli was supposed to be transformed to have the ability of antibiotic resistance. In the experiment E.coli was first transferred into both DNA- and DNA+ microcentrifuge tubes, and then manipulated into competency so that the plasmid can enter into the cells. E.coli is not naturally competent and therefore we tried to manipulate it by exposing it to abrupt heat and cold treatments and through the treatment of the metal cations of chloride salts. The samples were then transferred to Ampicillin positive and negative plates to test for growth. After incubation, if there was any growth on the plates, the plate was placed under a UV light to look for the green light from the protein. We hypothesized that about 10% of the e.coli would be transformed at the end of the experiment. We hypothesized that we would find bacteria most like the original non-transformed e.coli on the plates that did not contain ampicillin because it would not have acquire the ability to resist antibiotics. We also thought that we would find only genetically transformed bacteria cells on the plates containing