Title of Presentation or Performance
Expression and Isolation of the BCHE Protein from Dinoroseobacter Shibae in Rhodobacter Capsulatus
Submission Type
Event
Expected Graduation Date
2017
Location
Center for Natural Sciences, Illinois Wesleyan University
Start Date
4-16-2016 2:00 PM
End Date
4-16-2016 3:00 PM
Disciplines
Biology | Education
Abstract
Photosynthetic bacteria rely on photosynthesis to get energy. Bacteriochlorophylls harvest the light energy in photosynthesis and are important to these bacteria, but several enzymes used to synthesize this macromolecule are not completely understood. The gene bchE, from the marine bacterium Dinoroseobacter shibae, has been proposed to code for a protein used to help catalyze the formation of the bacteriochlorophyll structure. Isolation of polypeptide encoded by the bchE gene was performed to better understand this enzyme. Using conjugation, ST18 E. coli cells with plasmids containing the bchE gene were mated with a Rhodobacter capsulatus strain lacking functional BchE to allow expression. The Brp4 strains containing plasmids were grown to produce the BchE protein. GST tag column chromatography was used to isolate the BchE protein and associated proteins. Protein gel electrophoresis and Western blot techniques were used to ensure that the protein had been expressed in and isolated from the Rhodobacter capsulatus strain. The use of a photosynthetic bacterial host to express proteins is a new approach and will hopefully allow identification of proteins that interact with the BchE polypeptide.
Included in
Expression and Isolation of the BCHE Protein from Dinoroseobacter Shibae in Rhodobacter Capsulatus
Center for Natural Sciences, Illinois Wesleyan University
Photosynthetic bacteria rely on photosynthesis to get energy. Bacteriochlorophylls harvest the light energy in photosynthesis and are important to these bacteria, but several enzymes used to synthesize this macromolecule are not completely understood. The gene bchE, from the marine bacterium Dinoroseobacter shibae, has been proposed to code for a protein used to help catalyze the formation of the bacteriochlorophyll structure. Isolation of polypeptide encoded by the bchE gene was performed to better understand this enzyme. Using conjugation, ST18 E. coli cells with plasmids containing the bchE gene were mated with a Rhodobacter capsulatus strain lacking functional BchE to allow expression. The Brp4 strains containing plasmids were grown to produce the BchE protein. GST tag column chromatography was used to isolate the BchE protein and associated proteins. Protein gel electrophoresis and Western blot techniques were used to ensure that the protein had been expressed in and isolated from the Rhodobacter capsulatus strain. The use of a photosynthetic bacterial host to express proteins is a new approach and will hopefully allow identification of proteins that interact with the BchE polypeptide.