Jellyfish Green Fluorescent Protein Transformation via pGLO Plasmid (Lab Report Sample)
This is a lab report that required the customer/student perform genetic transformations on certain bacteria based on the gene that codes for green fluorescent protein (GFP) found in bioluminescent jelly fishes. The protein is the one that causes the jelly fish to fluoresce in the dark.
the lab involved transfering the gene from jelly fish cels onto bacterial cells via a pGLO plasmid, which is able to encode the the gene gor GFP onto another gene that confers the bacteria resistance to antibiotic ampicillin, as well as a gene regulation system which controls expression of the fluorescent protein in transformed cells.
My task was to study the experiment performed by the student and determine the degree of success in the efforts to genetically alter an organism (genetic engineering).
I was also to answer some questions provided by the professor on genetic engineering and biotechnology, and then write a lab report for the whole exercise.
pGLO Transformation Lab
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pGLO Transformation Lab
This Lab aimed at investigating the impact of pGLO plasmid on various E. coli colonies. PGLO is a genetically modified plasmid containing three genes. The first is the gene of resistance to ampicillin (antibiotic) that helps genetically engineered bacteria flourish in the presence of this antibiotic. The second gene is GFP which enables bacteria to fluoresce green light under the right conditions for the green fluorescent protein (UV-Light). The third ARA-C gene is a trigger for the activation of the GFP gene when the bacteria are exposed to arabinose sugar.
We used a transformed plasmid injected into E. coli in this experiment. Transformation is the way a plasmid is removed from the host and placed in a bacterial cell of another country. The bacteria will once in the new cell recognise the plasmid genes and express the desired phenotype. For this lab the transformed plasmid is inserted in E. with the aid of a process known as a heat shock by using a CaCl2 containing transformation solution. Bacteria were cultivated in four separate plates of growth to mimic various cultivating conditions. Ampicillin was not present on the first plate (-pGLO/LB) and bacteria lack the plasmid. As the bacteria will feed on the broth, a lot of development can take place in this bowl (LB). The colonies, however, do not fluoresce since a repressive protein will be attached as the working operon next to the promoter (starting point) of the GFP gene in this case, thereby enabling a stop of gene transcription because of lack of ARA. This illustrates the regulation of the GFP gene. The second plate (-pGLO / LB / AMP) does not expand because of the absence of pGLO plasmid, so the bacteria are not immune to ampicillin, and thus do not increase. The third platform (+pGLO/Lb/AMP) should display moderate growth, as the bacteria possess the plasmid that enables them to expand on ampicillin and feed on LB. The color of these cells should however remain white and due to the lack of sugar, the repressor will still be bound to the operator and thus the promoter transcription of the GFP gene was turned off. The fourth plate bacteria (+pGLO/LB/AMP/ARA) could glow because, with the ARA available, an ARA induction molecule
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