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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Bacterial Transformation Lab Report Backround: The plasmid pGLO contains an antibiotic-resistance gene‚ ampR‚ and the GFP gene is regulated by the control region of the ara operon. Ampicillin is an antibiotic that kills E. coli‚ so if E. coli‚ so if E. coli cells contain the ampicillin-resistance gene‚ the cells can survive exposure to ampicillin since the ampicillin-resistance gene encodes an enzyme that inactivates the antibiotic. Thus‚ transformed E. coli cells containing ampicillin-resistance
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The pGLO lab is a lab where students attempt to put the genes that make a jelly fish glow into E. Coli. After a process called transformation‚ the process in which a cell takes up and expresses a new piece of genetic information‚ the E. Coli will be able to glow and will be antibiotic resistant. The students first need to learn a couple of techniques before they are able to begin this lab. The first technique they will need is how to keep their environment sterile. They must learn to only open their
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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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Glowing Transformations Abstract In this experiment‚ the idea is to become familiar with the transformation of cells. A well thought out procedure‚ involving a heat shock procedure‚ a good antibiotic‚ an inducer known as arabinose to show the newly expressed DNA by a visible fluorescent glow‚ and a stable control group is what contributes to this experiments thoroughness. It is predicted that the four agar plates will all yield different forms of growth‚ with different coloration and colony
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