Gene Expression with E.Coli

Topics: Bacteria, DNA, Gene Pages: 9 (2012 words) Published: October 1, 2014
Title

Gene expression with E.coli bacteria through means of transformation with plasmid DNA

Abstract
Science has discovered that with gene expression and genetic engineering, DNA and organisms can be manipulated like never before. This has become an extraordinary discovery because it has lead us to countless medicinal products and cures for diseases and continues to serve as a great asset as research continues. This lab consisted of introducing a plasmid that contains an ampicillin-resistance gene into the common bacteria E.coli. With this, transformation is expected to occur and demonstrate how this prokaryotic, E.coli, uptakes a plasmid that allows it to become resistant to antibiotics and with this information, scientists will be able to use it for agricultures, vaccines, biodegradable products and so on. The experimental design allowed for the lab to be split up into two groups of plasmid. One of the groups had the “Control plasmid DNA” while the other had the “Plasmid lux DNA” and both group contained a No plasmid used as a negative control. A series of step such as ice baths and heat shocks were put into place to allow for transformation to occur and then the solutions were placed on an agar plate and incubated at 37° for a week. Our results failed to reject our hypothesis and predictions and thus resulted that there was indeed a high percentage of transformation efficiency. The agar plate with the ampicillin environment but no plasmid uptake resulted in no growth while the rest that were a variation between ampicillin environment and nutrient broth have both lawn growth and colonial growth.

Introduction
When talking about DNA and gene expression, let’s admit that there is tons to talk about. With proteins being the building blocks of life that are responsible for determining the chemical and physical structure of a cell or organism, it must be kept in mind that these molecules grabbed this information from somewhere else. This is where Gene expression comes into play. The basics of this process is having to do with DNA being converted to RNA through transcription and thus RNA converted to protein molecules through translation. Although this entire process is easier said than done, researchers describe Gene expression as extremely important in our biological standpoint. Tons of research is being done today through the manipulation of genes and the central dogma that could lead to the discovery and cure for many diseases that are incurable today. (Lorkowski et al. 2004) The technology in today’s date has advanced drastically that with the use of Biotechnology, scientists and researchers can modify and regulate gene expression the same way a cell can. Within Biotechnology, Gene engineering involved the direct manipulation of genes through techniques such as transformation and cloning. (Alberte et. Al 2012) E. coli is a prokaryotic, gram negative bacteria that is found in the digestive system. It is antibiotic resistant, meaning that throughout generations this organism has gone through mutations that allow it to be resistant to numerous medicinal products. Although this bacteria is not deadly, with only causing stomach pains and diarrhea, there are other bacteria more harmful that undergo the same process. This process is termed transformation. (Parker, 2003) In this laboratory, the objective was to introduce a plasmid (DNA) that contained an ampicillin-resistance gene into E.coli to demonstrate transformation. Ampicillin is an antibiotic introduced to treat certain infections caused by bacteria. (American Society of Health Pharmacists, 2010) With the use of various vectors and variables that are used to deliver foreign DNA into E.coli, the discoveries and conclusions will hopefully point the research in the right direction to allow for the future development of this process. Due to the low chances of transformation being successful, predictions involve...
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