Sulfolobus Oileatacus
Recombinant DNA in E. coli Expresses Devastator Enzymes to Clean Oil Spills
Dr. Land
University of the Pacific, Stockton
Abstract
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 …show more content…
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 the bacteria’s genome and therefore being translated into a …show more content…
The SmaI had cut the Sulfolobus Oileactacus at specific palindromes in the sequence of DNA and this isolates the devA gene from the rest of the plasmid. The microfuge tube was centrifuged briefly and placed in a 37°C water bath for ten minutes to allow the SmaI to digest the DNA, but no longer than 10 minutes otherwise the enzyme would begin randomly attacking the DNA. The tube was transferred to ice to stop the reaction from continuing. As seen in Figure 1 after digestion with the SmaI, the DNA fragments are of 1800bp and 600bp in
Dr. Land
University of the Pacific, Stockton
Abstract
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 …show more content…
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 the bacteria’s genome and therefore being translated into a …show more content…
The SmaI had cut the Sulfolobus Oileactacus at specific palindromes in the sequence of DNA and this isolates the devA gene from the rest of the plasmid. The microfuge tube was centrifuged briefly and placed in a 37°C water bath for ten minutes to allow the SmaI to digest the DNA, but no longer than 10 minutes otherwise the enzyme would begin randomly attacking the DNA. The tube was transferred to ice to stop the reaction from continuing. As seen in Figure 1 after digestion with the SmaI, the DNA fragments are of 1800bp and 600bp in