Molecular cloning and sequencing of penicillin G acylase from Shigella boydii

Abstract

In this study, 290 non-Escherichia coli Enterobacteriasea that were isolated from environmental and clinical specimen, were sent to the laboratory for examination with routine microbiological tests for identification of isolates. After identification, non-E. coli isolates were inspected by PCR for existence of penicillin G acylase (PGA) gene. Then, a PGA positive strain (Shigella boydii) from clinical specimens was selected for further analysis. First, DNA was isolated and PCR reactions were conducted using primers based on conserved region of PGA genes. The PCR reaction resulted in amplification of a specific product with expected length. The PCR product was cloned in pGEM-T Easy vector. Sequencing revealed that the gene, composed of encodes a polypeptide of 846 amino acid residues. Analysis of obtained sequence against databases showed the highest homology (about 96%) with the PGA gene reported from S. boydii.