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 pJET/bshA and pMG36e/bshA respectively. Both recombinants were transferred to E. coli MC1022 by chemical transformation. Obtained recombinants analyzed for expression and sequences. The results were confirmed that production of bile salt hydrolase from recombinant E. coli MC1022 pJET/bshA found to be higher while compared with E. coli MC1022 wild type and recombinant E. coli MC1022 pMG36e/bshA strain. However production of bile salt hydrolase from recombinant E. coli MC1022 pMG36e/bshA found to be higher while compared with E. coli MC1022 wild type. The recombinant plasmid also found to be stable in host organism after a few generations.