Pglo Plasmid on Genetic Transformation of E.Coli by Heat Shock

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  • Topic: Bacteria, DNA, Plasmid
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  • Published : April 30, 2013
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November 25, 2012
The Effect of the pGLO Plasmid on Genetic Transformation of E.coli by Heat Shock Introduction
Genetic transformation is the genetic alteration of a cell resulting from the direct uptake, incorporation and expression of exogenous genetic material l(exogenous DNA) from its surroundings and taken up through the cell membranes. This was first demonstrated in 1928 by Bacteriologist Frederick Griffith (Lederberg 2000).A plasmid is a small circular piece of DNA that contains important genetic information for the growth of bacteria. In a recent study, students tested the response of the arabinose operon promoter from E.coli to two different carbon sources. They directly observe the effects of these sugars on gene expression by monitoring the ability of the transformations to fluoresce under long-wave UV light. This experiment helped students experience transforming E.coli with plasmid DNA and using antibiotics for the positive selection of transformation (Mosher 2002). The present study was carried out in order to see if plasmids could be employed to transfer genes from one organism to another using a vector. The hypothesis was that the pGLO plasmid would introduce new genetic material to E.coli bacteria thereby changing the genetic make-up of the E.coli. Such a result would suggest that bacteria usually transfer these plasmids back and forth allowing them to share these beneficial genes. That is say, it is indispensable and important for pGLO plasmids to transfer GFP gene because a gene can be spliced into the DNA of a plasmid and when it is transferred from one bacterial cell to another, the gene will be transferred along with the plasmid DNA. Furthermore, we will also predict to find that the bacteria in non-transformation plate,which contains ampicillin, cannot survive. All bacteria which survive in the transformation plate with ampicillin must contain the ampicillin resistance gene and would fluoresce and glow in the dark. This...
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