Recombinant DNA in E. coli

Topics: DNA, Molecular biology, Polymerase chain reaction Pages: 4 (2774 words) Published: October 26, 2014

Recombinant DNA in E. coli Expresses Devastator Enzymes to Clean Oil Spills Dr. Land
University of the Pacific, Stockton
The bacteria Sulfolobus Oileatacus has an enzyme called the Devastator, DevA, that helps metabolize crude oil. Since the bacterium is a thermophile, it won’t survive at normal temperatures and cannot be used to clean up oil spills. The devA gene was cloned through the polymerase chain reaction and was confirmed to be present using a restriction enzyme digest; after which the cloned genes were inserted into the bacteria E. coli through the transformation process consisting of a restriction enzyme digest and ligation. Colonies of the bacteria were grown; plasmids from each colony were purified, and then analyzed using the agarose gel electrophoresis. Results indicated that only the bacteria that had the plasmid with the correctly oriented devA gene inserted would be able to metabolize crude oil as an energy source. This experiment is important because E. coli that expresses the devA gene can be used to clean up oil spills at normal temperatures. Further research could be done on the transformation of bacteria and use of them in the environmental field. Introduction

All species on earth have DNA and differ only in the sequence of the DNA nucleotides. Because of this similarity shared by organisms, DNA can be manipulated and inserted into the genome of other organisms. This process is called DNA cloning which replicates a gene of interest using restriction digests and the polymerase chain reaction. The specific cuts in the DNA sequence that restriction enzymes make and the joining of DNA together by the enzyme ligase allow for the production of millions of copies of the gene of interest. In the context of bacteria, heterologous protein expression occurs when a protein is expressed in bacteria which normally do not make that particular protein (4). The expression comes from the gene encoding for the protein being inserted into...

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