Pglo Lab

Introduction: Transforming a gene or genetic information from one organism into another with the hopes that if done successfully the organism with the new DNA will be given new traits is a method known as genetic transformation (Rafter). Genetic transformation is used quite frequently in today’s world, form medicine to agriculture.
In this lab we will be inserting a gene into an Escherichia coli bacteria with the help of a plasmid. Escherichia coli bacteria also known as E. coli, is a bacterium that is rod shaped and contains flagella to help it move. The bacterial DNA is circular inside of an E. coli bacterium. E coli. is most known for being found in the intestine of humans and animals but it can also be found in other places such as food
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coli bacteria new traits. The pGLO plasmid that is being transformed into these cells contains genes that can give colonies of bacteria the ability of antibiotic resistance and a green fluorescent glow. Four different models were prepared and plated on multiple agar plate. After the bacteria was grown for three days in an incubator at 37°C; observations were made and recorded (Table 1). All of the plates were looked at for the amount of colonies grown, if growth was present, and if the colonies gained the ability to glow green. Transformation was successful in the plates where the bacteria consumed the pGLO plasmid. In the first plate that the bacterium was plated on it included the LB broth and of ampicillin antibiotic (amp), 2 colonies were present. The second plate of bacteria was grown with the presence of LB broth, ampicillin, arabinose sugar (ara), and 22 colonies were observed. But a green fluorescent glow of the colonies was only present in plate 2. Plates 3 and 4 were the control plates. The bacteria were not transformed and no growth was present in either
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Our hypothesis was that if the plate contains only the LB broth the E. coli bacteria would have no antibiotic resistance and would not glow. If the plate contains just LB broth and ampicillin then the E. coli bacteria will have antibiotic resistance only if the plasmid is present. If the plate contains LB broth, ampicillin and arabinose then the E. coli bacteria will glow fluorescent under a UV light and it will have antibiotic resistance. Similar to our expectations our results suggested that our hypothesis was correct. This is due to the fact that n order for the E coli. bacteria to gain either of the traits the plasmid had to be present in the first place, because the GFP gene is inside the plasmid. If arabinose is not in the media in which the bacteria was growing on then the GFP gene could not turn on, thus the bacteria can not glow. This is why the LB/amp/ara plate was the only one to express both traits(antibiotic resistance and glowing). The second part to our hypothesis was that using HIC,GFP would be purified and the final tube would express GFP under an ultra violet light. Contrary to what we expected the final tube nor any of the columns expressed the ability to glow under ultra violet light. This unexpected result may have been