Pglo Transformation Lab Essay

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. Our hypothesis was that the transformed solution with no plasmid DNA and ampicillin would produce no bacteria colonies, as it wouldn't be able to grow without the gene for ampicillin resistance. Also, the transformed solution with just LB and ampicillin would produce bacteria colonies but the transformed solution with LB/ampicillin/Arabinose would produce glowing bacteria colonies (as Arabinose allows the GFP gene to be expressed, but in both cases bacteria colonies would be present because of the gene of resistance to the antibiotic, ampicillin).

We essentially made the required transformed solutions--and the controls--swiped them on the agar plate, and then observed to see whether or not bacteria colonies grew and whether or not they glowed. Our data fully supported our hypothesis. We can thus conclude that bacteria can take in foreign DNA through the process of transformation and that this foreign DNA can fundamentally change the bacteria (ex: making it glow). Future research can involve inserting other pieces of DNA into bacteria from different organisms, making the bacteria take on various phenotypic characteristics.

Background Information:

Genetic transformation is one of the most important processes in biotechnology. Essentially, genetic transformation involves the process where a cell (in this lab, a bacterial cell) takes up foreign DNA from its surroundings and incorporates it into its own DNA. This gene transfer is accomplished with the aid of a plasmid, a naturally-occuring...


2identified that the presence of Arabinose codes for the protein necessary to create the glowing effect, the transformation was successful through confirmations through the results. Introduction The insertion of a gene into an organism is a process known as bacterial transformation, this is used to alter the trait of an organism. Bacteria will incorporate new DNA into it’s own DNA. Bacterial Transformation is used often by scientists in order to benefit society, bacterial vaccine’s can be transformed from bacteria in order to prevent infection and disease in animals. Scientists study the numerical and analytical methods of genetic transformations and the mutations that may occur with these changes, they then apply this to their experiments (Redfield, R., M. Schrag, and A. Dean, 1997). A plasmid, which is a self-replicating DNA molecule separate from it’s chromosome, was used in this experiment to help transform. The plasmid, PGLO, contains the genes that codes for green fluorescent protein (GFP). Green fluorescent protein was originally found in the jellyfish, Aequorea Victoria.It also carries the gene which resists antibiotic ampicillin (bla gene) and one that codes for the arabinose C protein (araC). This plasmid was chosen for this specific experiment because if the transformation is successful the organism will glow a fluorescent green when exposed to ultra violet light. The process of heat shock was used to change the fluidity of the membrane, without this step DNA would not be able to enter the bacteria at a productive rate. The hypothesis stated was that if the GFP gene is placed in E.coli it will glow, therefore making the transformation successful. Materials & Methods The steps to conduct this experiment must be done with extreme caution in order ensure accuracy. Two micro test tubes must be labelled, ensuring the caps are closed, one of them

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