Workshop: pGLO Bacterial Transformation                               By Belinda Román and Ricardo Chiesa         Bacteria...
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pGLO Bacterial Transformation and pGLO SDS Page Extension

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Transcript of "pGLO Bacterial Transformation and pGLO SDS Page Extension "

  1. 1. Workshop: pGLO Bacterial Transformation By Belinda Román and Ricardo Chiesa Bacterial transformation involves the process of inserting a certain gene from oneorganism into another organism. The purpose of this is to make an organism express a gene thatis not present in its own genome. Scientists use different techniques to do so, and one of them isthe use of a plasmid. Plasmids are pieces of an autonomous replicating DNA obtained frombacteria. Genes of interest are inserted into plasmids, and then by introducing plasmids intobacteria, transformation will occur As a result, the host organisms will express the proteins ofinterest. In the lab, we worked with this process, but we used Green Fluorescent Protein (GFP) asa biological marker to see if bacterial transformation occurs. First of all, we make cultures of E.coli bacteria in Petri dishes. One of the cultures had the plasmid that was going to transform thebacteria. The following week, we looked at this culture with UV light, and transformation didoccur because GFP was expressed in E. coli. Afterwards, we practiced the technique of SDS-PAGEto divide and observe proteins of the transformed bacteria in a polyacrylamide gel. Finally, weobserve the proteins with UV light and Coomassie staining to see the location of GFP. Our resultsshow that partially denatured proteins present a band of 37kD, and the completely denaturedproteins show a band of 27kD.

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