Lb-Pglo Lab Report
n this lab, there were four objectives needed to be met. The first one was to perform a genetic transformation procedure, the second was to move genes from one organism to another using a plasmid as a vector, and the third was to manipulate tools of biotechnology. The bacteria E. coli was used to manipulate and transform. The E. coli would be tested for ampicillin resistance and a green fluorescent glow. One hypothesis made for this lab was that the bacteria that developed a resistance to ampicillin would reproduce even in the presence of the ampicillin. Another hypothesis made was that the bacteria would glow with the addition of the sugar arabinose.
All three of the objectives seem to go hand in hand. The lab began by inserting transformation
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The plate labeled LB: -pGlo had a lawn of bacteria, which meant that all of the bacteria that reproduced was able to survive. The plate labeled LB/amp: -pGlo had no bacteria present at all. This could mean that the -pGlo, the pGlo that received none of the plasmid DNA, had no resistance to ampicillin which would lead to the conclusion that normally, E. coli is not resistant to ampicillin. The plate labeled LB/amp had several colonies of bacteria; this meant that the bacteria that contained the plasmid DNA was able to survive and replicate in the presence of ampicillin. The plasmid DNA is the source of the resistance to ampicillin because that is the only thing different between the LB/amp: -pGlo, which had no bacterial reproduction and the LB/amp: +pGlo, which had a large amount of bacteria survive and reproduce. The plate labeled LB/amp/ara: +pGlo had several surviving colonies like the other +pGlo plate, except this time the colonies were a green color and the glowed under a UV light. The glow would be a result of adding the sugar arabinose, which seems to be acting as an inducer in the operon of the bacterium. It can be considered an inducer because in the presence of the arabinose, the colonies glowed, but without it the colonies did not experience any change, leading to the belief that the gene that leads to the green fluorescence is normally turned off.
To further the research on the effect of the arabinose on the E. coli, a sample of the bacteria in the LB/amp: +pGlo and placed it in a plate with arabinose lines across the bottom. This plate was allowed to incubate overnight, When the plate was revisited, and found that the colonies of bacteria were a green color. When observed under a UV light, the bacteria glowed. This proved that arabinose is the cause of the green
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