Cloning and overexpression of an E. coli pbp1a

CHED 202

Courtney M. Alexander, Lilian T. Lamech, and Kimberly M. Specht, spechtk@denison.edu. Department of Chemistry and Biochemistry, Denison University, Ebaugh Laboratories, Granville, OH 43023
The periplasm of the bacterial cell wall contains the peptidoglycan layer. Peptidoglycan is a mesh of cross-linked sugars and peptides that is responsible for maintaining cell shape and integrity. The peptidoglycan is synthesized by bifunctional enzymes with transglycosylase and transpeptidase activity known as penicillin-binding proteins (PBPs). We will describe the cloning and overexpression of a putative PBP from Burkholderia cenocepacia. The genome for this strain,J2315, has been sequenced at the Sanger Institute. The open reading frame identified as BCAM 2632 has been identified as a gene homologous to PBP1a from E.coli. Studies characterizing the BCAM 2632 gene product will be described.