Cloning and Characterization of Gene Promoters from Bacillus pumilus

DNA fragments obtained from Sau 3AI partially digested total DNA of Bacillus pumilus UN31 C 42 are first inserted into Bam HI site of pSUPV4, a promoter probe vector. The recombinant DNA molecules are transformed into Escherichia coli cells and eight three Kan r 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.