The bile salt hydrolase gene (bshA), encoding bile salt hydrolase enzyme (EC 3.5.1.24) from probiotic isolate Lactobacillus acidophilus Ar strain which is responsible for assimilation cholesterol were studied in the present work. About 801 bp in length DNA fragment of Lb. acidophilus Ar strain was amplified by PCR techniques. Two restriction sites (PstI/SacI) were added to each end of that fragment for manipulation of DNA during cloning. Amplified fragment inserted into pJET1.2\blunt end vector and pMG36e vector respectively. pJET1.2\blunt end vector is overexpression plasmid for E. coli MC1022, and pMG36e vector is a shuttle vector which is able to replicate in both E. coli and lactic acid bacteria. The resulted constructs were named as pJ

Recently, gallbladder stones have been contained bile salt saturated a proximal 70 % cholesterol. This led us to investigate how can use transformer Streptococcus salivarius with plasmid pMG36bsh to fragment cholesterol of gallstones in vitro. Total mRNA of S. salivarius was produced using easy-spinTM, total RNA extraction kit and PCR cDNA-RT to observe the change after percent pMG36bsh vector and prepare S. salivarius have two copies from bsh genes (cgh, bsh) to fragment gallstone in bacterial culture. Our data shows increase bacterial bsh expression help to reduce gallstones concentration in culture when bile salt presented as stimulating agent for the association bsh genes were 77% compare with wild type has the reducing concentration ra