understand the process of transformation and the effects on gene expression. The pGLO(-) culture had growth on the LB medium‚ while the LB amp and LB amp + ara mediums had no growth. It was expected that the LB medium had growth on the plate because it served as a control. The LB amp and LB amp + ara had no growth or glow under UV light because they were not successfully transformed and still contained the antibiotic ampicillin that prevented the growth of E. coli (3). The pGLO(+) culture had growth
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Transformation Of Escherichia Coli With pGLO Plasmid April 24‚ 2013 ABSTRACT: This experiment focuses on genetic engineering and transformation of bacteria. The characteristics of bacteria are altered from an external source to allow them to express a new trait‚ in this case antibiotic resistance. In is experiment foreign DNA is inserted into Escherichia coli in order to alter its phenotype. The goal of the experiment is to transform E. coli with pGLO plasmid‚ which carries a gene for ampicillin
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Transformation in bacteria is the genotype alteration by the uptake of naked‚ foreign DNA from the environment. This concept of transformation was first discovered when Fred Griffith an experiment using mice and strains of pneumonia. Griffith concluded that a “principle” was transferred from heat-killed S strains to the R strains‚ which transformed them into deadly S strains. Oswald Avery later determined‚ through a series of experiments‚ that DNA was the “principle” that caused the R stains to become
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Genetic Transformation of Bacteria Abstract The transformation of bacteria was successfully carried out using a plasmid carrying a gene that codes for green fluorescent protein‚ which gives a signature green glow reminiscent of a jellyfish. This gene‚ however‚ is only active when the sugar arabinose is present. A gene coding for antibiotic resistance was also found within the plasmid and served as a means to verify that transformation had indeed taken place. The hypothesis was that the bacteria
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The pGLO plasmid‚ which has the bla and GFP gene‚ was developed so it can operate like the arabinose operon and be able to transcribe genes in the presence of arabinose sugar. In this plasmid‚ the cluster of genes is regulated by the spontaneous on/off element by a single promoter‚ which is dependent on the DNA binding protein araC. This protein is at the binding site for RNA polymerase at the beginning of the operon‚ so when arabinose is present (like in one of the experimental plates)‚ it is taken
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Genetic Transformation with pGLO Plasmid Introduction: Genetic transformation is where one organism takes on a characteristic from another organism (Bacterial Transformation 2013). For this experiment we used the bacteria E. Coli to take in foreign jellyfish DNA which will allow it to change genetic material. This experiment determines the effects that the plasmid pGLO has in transferring the Green Florescent Protein found in a jellyfish into the bacteria. It determines whether or not pGLO acts
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Introduction Bacterial transformation is the permanent alteration of a bacterial cell genotype as a result of its uptake and incorporation of foreign DNA fragments from external medium (Anthony et al‚ 2008). In addition to chromosome‚ bacterial cells often contain extrachromosomal DNA called plasmids which are capable of autonomic replication and antibiotic resistance (Dale & Simon‚ 2010). Plasmids can transport foreign DNA into host or other bacterial cells hence they are known as vectors.
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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
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Abstract: The topic of this research involved the occurrence of genetic transformation in bacteria (E. Coli). More specifically‚ a previously prepared pGLO plasmid--which consisted of the gene to be cloned--was used to transform non-pathogenic bacteria. The pGLO plasmid contained a gene for the Green Fluorescent Protein (GFP) from a bioluminescent jellyfish and a gene for resistance to ampicillin‚ an antibiotic. Essentially‚ we wanted to determine the conditions of the bacteria that would glow.
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Genetic transformation of Escherichia coli with pGLO (Adapted from: Biotechnology Explorer: Bacterial Transformation: The pGLO System. Instructors Guide. BIO-RAD). Objectives a. To understand one of the most commonly used techniques for introducing DNA into E. coli cells and its use in molecular cloning. b. To become familiar with the concept of using green fluorescent protein (GFP) as a molecular tag for studying gene expression in bacteria and other organisms.
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