Pglo Transformation Lab Report

Transformation of Escherichia coli.

Introduction The purpose of this lab is to genetically transform the bacteria Escherichia coli using plasmids. Transformation is the process of inserting recombinant DNA into cells (Hillis, 2019, concept 13.2). Bacterial transformation is commonly used to make insulin, and to produce disease resistant crops. The purpose of this transformation is to add the gene Green Fluorescent Protein (GFP) into bacteria. Transformation is achieved by transforming the plasmid pGLO into the bacteria. pGLO has been inserted with GFP, a gene that is resistant to the antibiotic ampicillin. GFP is a gene found commonly in the species Aequorea victoria, which is a bioluminescent jellyfish. In order to isolate GFP to insert it

In this lab, antibiotic resistance is demonstrated when pGLO DNA is able to grow on the agar plates with ampicillin. Plasmids are able to incorporate genes into their DNA because of bacterial competence. Bacterial competence is the ability of bacteria to take in any stray DNA that they encounter. This ability allows the plasmid to integrate GFP and the gene for

When the bacteria integrates GFP into its DNA, it allows the bacteria to glow in the dark. The ability of GFP bacteria to glow in the dark is controlled by an operon. An operon is a genetic regulatory system that can be found in bacteria (Britanica, 2018, “operons”). An operon includes an operator, which acts as a switch that turns genes off and on. Operons can be inducible or repressible. The GFP operon is inducible (Mcauley, 2021, pGLO). This means that in the presence of an inducer, the operon will produce GFP. The inducer that causes GFP to form is arabinose. Repressible operons are the opposite of this. When the repressor is present, it will turn off the formation of the product. Hypothesis If E coli bacteria is added to all four agar plates, then both of the agar plates labeled +pGLO and the -pGLO plate without ampicillin would grow. Variables The independent variables in this experiment is the addition of pGLO, ampicillin, and arabinose. The dependent variables in the experiment are whether or not bacterial growth occurs, or if the bacteria grow. The constants in the experiment are the size of the agar plate, the temperature in which the agar plates incubate, and the length of time the