Purification and Characterization of BaeI and PksL in Bacillus amyloliquefaciens and Analysis of Protein Activity within the Bacillaene and Difficidin clusters of Bacillus Subtilis

UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)
Alexis Lewars (Creator)
The University of North Carolina at Greensboro (UNCG )
Web Site: http://library.uncg.edu/
Jason Reddick

Abstract: In these experiments, the genes PksL and BaeI were purified and cloned into vectors such as Top10 and BL21 star. SDS-Page and Gel electrophoresis were used to confirm the transformation of these genes. The sequence of BaeI was confirmed through Sanger sequencing. A reaction of PksL with CoA and Sfp was run to determine the successful addition of the phosphopantetheine arm to the acyl carrier protein. Bacillaene and Difficidin were assigned homologs by structural and genetic analysis using protein sequencing provided by NCBI. The sequences of such amino acids were analyzed for distinct characteristics. Future experiments within the Reddick laboratory will involve reactions with BaeI and PksL to determine double bond placement within Difficidin polyketide synthesis.

Additional Information

Honors Project
Language: English
Date: 2020
Polyketide Synthesis, BaeI, PksL, Proteins, Biochemistry, Antibiotics, Protein, Antibiotic, PksX, Curacin, Difficidin, Bacillaene, Homolog, Mupirocin, Oocydin A, Pederin, Diaphorin, Psymberin, Onnamide, Polyketide, Beta-Branching

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