Plasmid, a pGFPuv developed by Clontech, contains a gene for the green fluorescent protein (GFP) that comes from the bioluminescent jellyfish Aequorea victoria. This gene allows the jellyfish to produce a protein that fluoresces, or glows, under ultraviolet light. pGFPuv is used as a vector to transform E. coli bacteria. The GFP gene of pGFPuv is switched on in the presence of lactose.
The plasmid also contains a gene for ampicillin (antibiotic) resistance. Cells with Ampr produce a protein that decomposes ampicillin and enables them to survive in an ampicillin environment.
Perform the following steps in a laboratory:
Introduce the plasmid pGFPuv in E. coli cells by subjecting them to heat shock in the presence of calcium chloride.
Spread samples of these cells on four sterile agar plates. Three plates should contain nutrient agar (NA) and one should contain Luria Bertani agar (LB).
NOTE: LB agar contains lactose. NA contains glucose.
Based on the treatment meted out in each plate, write down your observations and conclusions. Compare the results with your peers.
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