cloning of bacillus subtilis phytase gene construct in escherichia coli

Background and objectives: phytase has a hydrolysis function of phytic acid, which yields inorganic phosphate. bacillus species can produce thermostable alkaline phytase. the aim of this study was to isolate and clone a phytase gene (phy) from bacillus subtilis in escherichia coli. materials and methods: in this study, the extracellular phyc gene was isolated from bacillus subtilis phytase c. after purification of the bands, dna fragment of phy gene was cloned by t/a cloning technique, and the clone was transformed into escherichia coli. afterward, the pgem-phy was transferred into e. coli top-10 strain and the recombinants were plated on lb agar containing 100 μg/ml ampicillin. the colonization of 1171 bp of gene phytase c was confirmed by pcr. the presence of gene-targeting in vector was confirmed with enzymatic digestion by xhoi and xbai restriction enzymes. results: the phytase gene was successfully cloned in e. coli. the result of cloning of 1171 bp phytase gene was confirmed by pcr assay. conclusion: our impression of this article is that several methods, such as using along with microbial, plant phytase reproduction, or low-phytic acid corn may be the better way from a single phytase.