transformation of E. coli bacteria with a plasmid that codes for Green Fluorescent Protein (GFP) and contains a gene regulatory system that confers ampicillin resistance. A plasmid is a genetic structure in a cell that can replicate independently of chromosomes. In this lab‚ the Green Fluorescent Protein‚ which is typically found in the bioluminescent jellyfish Aequorea Victoria‚ was cloned‚ purified‚ and moved from one organism to another with the use of pGlo plasmids. It was hypothesized that if bacteria
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transformation effici ency‚ with a higher ratio leading to higher transformation efficiencies. In order to asse ss this model‚ we transformed wild type Escherichia coli (BW25113) and a smaller mutant strain (JW2500) with different amounts of the plasmid vector pUC19.The mutant strain JW2500 with a lower SA:V ratio but a similar surface area to the parent strain displayed 70% ± 5% of the number of transformants from the w ild-type strain BW25113. Suc h results suggest a direct relationship
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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
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In this lab‚ pVIB plasmid will be used. A plasmid is a segment of DNA that can incorporate itself into the bacterial DNA. Although is not required for growth of the bacterial cell‚ plasmids can provide advantages in stressful environments such as the ability to adapt as environmental changes occur. In this lab‚ we will obtain a better understanding of bacterial transformations using pVIB. Hypothesis: If the bacteria transforms successfully with the pVIB‚ the the +plasmid (LB & AMP) will
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Abstract The purpose of this experiment was to study the transfer of genetic information on plasmid F’lac by using Escherichia coli. Plasmid transfer was measured by using two different methods. The first one was by using selection and contraselection with three antibiotics: streptomycin(which was replaced by naladixic acid for the second part of the experiment)‚ampicillin and kanamycin and the second one by using a colour indicator ( X-gal). As significant results‚ the percentage of transfer for
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Transformation of E. coli by plasmid DNA 1. Table showing the results from the selective plates |Plate |Plasmid |Contents of plates |Number of colony | | | | |White |Blue | |1 |Ligation mixture |Ampiclillin + X- gal + IPTG |10 |0
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This lab works to develop the understanding of bacterial transformation through the integration of a plasmid into E-coli bacteria. Understanding of plasmids‚ GFP‚ expression of genes and bacterial candidates is used to formulate a lab which demonstrates a variety of factors associated with transformation efficiency. It was deduced that there are certain requirements present in the pGLO plasmid for full gene expression and that an increase in transformation solution positively impacts the results
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isolated then treated with Sal I restriction enzyme to generate inserts (smaller fragments of DNA). Sal I restriction enzyme was also used to treat the vector plasmid in order to digest the V. fischeri DNA fragments. The inserts and the vector were then ligated together. E.Coli cells were then made competent in order to take up the plasmid DNA by transforming these competent cells with a “ligation mixture to create a population of host bacteria containing different combinations of the ligated inserts
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fischeri DNA and pGEM DNA was cut with restriction enzyme Sal I to prepare both for ligation. 3. Vibrio fischeri and pGEM DNA were ligated with T4 ligase. 4. E.coli DH5α competent cells were prepared for us so that transformation of the recombinant plasmids and the competent cells through heat shock transformation. 5. The transformation was then plated on LB/amp/X-gal plates. 6. The plates were screened for our target with ampicillin resistance‚ blue/white colony‚ then bioluminescence. 7. A bioluminescent
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Introduction In this week’s laboratory period students had the opportunity to perform a common procedure preformed by many if not all microbiologists known as genetic transformation. Genetic transformation is the ability to move DNA into an organism and thereby altering its genotypic and genetic makeup (2). Genetic transformation has shown to have a wide variety of uses in many scientific studies. In agriculture‚ gene coding for traits such as frost‚ pest‚ or spoilage resistance have been genetically
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