Formal Report - Bacterial Transformation

Formal Report - Bacterial Transformation - Escherichia Coli...

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Escherichia Coli Growth after Exposure to Various Plasmids Anil Kanungo BIO 206, Spring, 2007 April 5, 2007 1
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Escherichia Coli Growth after Exposure to Various Plasmids Abstract: Escherichia coli are a highly versatile bacterium which scientists have used to study genetics. Plasmids may easily be used to regulate gene expression in bacterial cells and were used in this experiment. For this experiment, culture plates were injected with 25 μL of a plasmid or control solution and E. coli cells. Half the culture plates contained ampicillin, which is known to prevent E. coli growth, and the other half did not. After incubating plates for a week, observations were made on whether the plasmids affected gene expression as expected. The observations made shed light on the general field of genetic engineering in microorganisms. 2
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Introduction: Plasmids are DNA molecules that are separate from the chromosomal DNA. They are double-stranded, circular, and capable of autonomous replication (Wikipedia: Plasmids 2007). Every plasmid has one DNA sequence that serves as an origin of replication which enables the plasmid DNA to be duplicated independently from the chromosomal DNA (Wikipedia: Plasmids 2007). Because plasmids are easily manipulated and can be used to easily isolate specific sequences from the rest of the genome, they were ideal for this laboratory experiment (Casali 156). The discovery of plasmids opened a door in the field of genetic engineering. Genetic engineering has allowed for “the correction of hereditary defects, the improvement of crop plants and domestic animals, and the production of proteins by microorganisms” (Lab Manual 8). Escherichia coli have been one of the most commonly used organisms in microbiology (Vaillancourt 32). According to Wikipedia, “its structure is clear, and it makes for an excellent target for novice, intermediate, and advanced students of the life sciences.” The objective of this laboratory experiment was to observe E. coli and see if it was possible to alter the DNA sequences being expressed by conjugating plasmids. Different plasmids were used to express features such as bioluminescence and melanin production. A control solution was also used to observe normal E. coli growth. Because of the proficiency of E. coli and the simplicity of plasmids, they were perfect candidates for observation. Materials and Methods: To begin this experiment, each lab group first cleaned off their work area with ethanol and paper towels. An ice box containing E. Coli cells, plasmid solutions, and a control solution was then obtained from the lab TA. The plasmid/control solutions were labeled C, P1, P2, and P3.
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