Cloning and Characterization of Gene Promoters from Bacillus pumilus

DNA fragments obtained from Sau3AI partially digested total DNA of Bacillus pumilus UN31-C-42 are first inserted into BamHI site of pSUPV4, a promoter-probe vector. The recombinant DNA molecules are transformed into Escherichia coli cells and eight-three Kanr clones (named pSUBp1- pSUBp83) are obtained. The inserted fragments in pSUBp53, pSUBp57, pSUBp21, which showed high level of kanamycin - resistance, are sequenced and analyzed, respectively. These fragments contain some conserved sequences of prokaryotic gene promoters, such as TATAAT and TTGACA box. The promoter fragment Bp53 could efficiently promote the alkaline protease gene of B.pumilus expression not only in E.coli but also in B.subtilis cells.