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The Process Of Inserting The Gfp – Containing Plasmid Of P Glo Into E. Coli By Means Of Heat Shock Transformation

1984 words - 8 pages

Introduction
The history of genetic transformation dates as far back as 1952 with Joshua Lederberg (Grote 2008). At this point not much was known about genetics; therefore what we now know as the episome was originally named a plasmid by Joshua Lederberg. The Current model of the plasmid was not found until the 1970’s. The plasmid then became the foundation for the rest of recombinant DNA technologies. Taking the form of a ring of DNA, the plasmid is passed down to the offspring in a hereditary like fashion. This means that bacteria, one of the organisms that use plasmids, will not reject plasmids but instead accept them into their cells and begin using them for protein synthesis and DNA replication. This increased knowledge gave us the ability to replicate specific sections of DNA. This was shown in a recent study in which DNA fragments were inserted into a plasmid and that plasmid then inserted into a bacterium (Cohen 2013). The DNA was then later seen being represented by the bacterium. This research was carried farther and included a similar result with full DNA from a different species of bacteria. Another study was done that indicated that plasmids were able to contain information that affects other things that enter the bacterium. These can either be other DNA molecules or it could come in the form of another vector (Gognies et al. 2012). Other methods are being researched into that can use larger segments of DNA that would not be plausible for a plasmid (Miyazaki and Roelof 2013).
Genes can be transferred in many unique ways each with a different rate of success efficiency (Weedman D 2013). The three most common are currently projectile bombardment, electroporation, and heat shock. The method of transformation that was used in this lab was heat shock. With this method the membrane of the cells became more permeable due to an increase in temperature. This sharp increase followed a gradual decrease in temperature to make the effect of the heating more effective. Plasmids that were in the surrounding area were then able to pass through the cell membrane and became a part of the cell its self.
In this experiment we inserted a plasmid into E. coli bacteria of which when in contact with a sugar named arabinose would glow under a florescent light. This is significant because it will show that DNA can be copied and represented in the genes when inserted through a plasmid. This is including the fact that the gene its self is not included in E. coli’s initial genome. It is also significant because the plasmid contains a gene that produces an ampicillin resistor. This will allow the bacteria to grow even in an environment that contains that antibiotic. My hypothesis was that the bacteria that had the plasmid that included the genetic material for pGLO and was cultured on the plate that had ampicillin and arabinose grew and when placed under a florescent light glowed a green florescent color. This bacterium also grew in a...

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