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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In this particular situation we didn’t add enough PGLO into the DNA so ours didn’t glow. In the control lab a different outcomes was observed in each of the four plates. In the LB/amp/arabinose agarose plate containing the +pGLO sample‚ fluorescent green colonies developed. This is because the gene which codes for the fluorescent protein‚ GFP‚ is located near the beta lactamase gene on the pGLO plasmid‚ which protects bacteria from the antibiotic ampicillin. When the cell produced beta lactamase
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bacterium integrates a piece of DNA into its genome‚ bacterial transformation has occurred. In this experiment bacterial transformation will be done using calcium chloride/heat shock. This is done by incorporating the plasmids into chemically competent cells that were made permeable by the calcium chloride solution and heat shock. In 1928‚ Frederick Griffith‚ a physician from London‚ was he first person to experiment with bacterial transformation. He permanently transformed a safe‚ nonpathogenic bacterial
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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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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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Student Lab Section 6 E. Coli 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
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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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Lab Report (Scientific Paper) 2: Bacterial Transformation;DNA Extraction Part I & II:Total Genomic Extraction & Plasmid Extraction;Electrophoresis By:Chris Foster Abstract: We conducted three experiments that included a Bacterial Transformation‚ a two process DNA extraction‚ and a final procedure using gel electrophoresis. The Bacterial Transformation lab was performed to prepare the plasmid into a bacteria and to use that bacteria to amplify the plasmid in order to make large quantities
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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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