Transformation and over expression of chromosomal bile salt hydrolase gene A (bshA) that transfer from transformer Escherichia coli to Enterococcus faecalis

Abstract

The clone Escherichia coli MC1022 as replication host carrying plasmid pMG36e (4.5 kilobases) which confer erythromycin resistance was used in this study as a vector that have a clone of chromosomal bile salt hydrolase gene A (bshA) of Lactobacillus acidophilus Ar and prepare for transfer to stereptococcus species. A 801 bp bile salt hydrolase gene A fragment cloning for hypercholesteremic treatment in human blood by bacterial biodrug when gene overexpression occurrence. The recombinant plasmid pMG36e called pMG36/bshA vector was extracted from E. coli using ethanol 96% precipitation method. Natural transformation method was used to transform Enterococcus faecalis, which supplied by Symbiopharm company as probiotic supplement. Detection of cloning gene depend on erythromycin resistance character and overexpression assay of bile salt hydrolase enzyme using specific activity of bile salt hydrolase enzyme in transformer strain of E. faecalis and compared with wild type of E. Faecalis, the enzyme activity was increased from (397 to 607) U\mg.