Genetic Transformation of Escherichia Coli with Pglo

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Genetic Transformation of Escherichia coli with pGLO Ahmed Islam Abstract Aim: This experiment is designed to help understand the concept of genetic transformation. This is the uptake of DNA fragments from the environment by a competent bacterium. Competency must be induced in bacterium such as Escheria coli. Also, this lab helps understand the concepts of plasmids, specifically pGLO, and their genes, specifically green fluorescent gene (GFP). Expression will be regulated using promoters. This lab deals with the promoter known as araC, and it plays an important role in the expression of fluorescence. Method and Materials: Competency was induced in E. coli, using calcium chloride. The pGLO was then introduced to the E. coli, and the mixture was plated onto four plates. 2 plates were plated with non-transformed E. coli the plates were designed to either promote or inhibit two factors: growth and fluorescence. The transformation efficiency was then calculated. Results: Results went as expected. Non-transformed E. coli did not grow in the presence ampicillin nor was there any fluorescence. All plates containing pGLO transformed E. coli had growth. Fluorescence was found only on the plate containing LB agar, ampicillin, and arabinose. The transformation efficiency was calculated to be 7219 transformants/ µg of pGLO DNA. Conclusion: there was some variability in the number of transformants/µg of pGLO DNA. This can be due to number of things. Timing and aseptic procedure are two main causes of variability. Introduction Genetic Transformation is a type of genetic recombination that allows competent bacteria cells to pick up DNA fragments from the environment and express the genes in the fragments (Kovach, 2012). In 1928, a British Medical Officer by the name of Fredrick Griffith published a report titled “The Significance of Pneumococcal Types.”

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