[Construction of promoter probe vector pFDC4 and gene expression vector pFDC11 with high transformation efficiency in...

A 0.5kb fragment from Sau3A-digested total DNA of Bacillus stearothermophilus CU21 was cloned with the vector pPGV5, a derivative of pPL703. The insertion of this fragment can activate the expression of the promoteless cat-86 gene on the cloning vector in both B. stearothermophilus and Bacillus subtilis hosts. When the 0.54 kb fragment is present in pPGV5 in either orientation, the transformation efficiency of the plasmid is increased about 10(3) to 10(4) fold in CU21 protoplasts. Southern hybridization showed this 0.54 kb fragment was homologous with a 1.6kb fragment, which was shown by Imanaka et al. (1984, J. Gen. Microbiol. 130, 1399-1408) to originate in a cryptic plasmid resident in CU21 and to enhance the transformat on efficiency of another plasmid. With this 0.54kb fragment a new promoter probe vector pFDC4 and a gene expression vector pFDC11 were constructed. Both can transform the CU21 recipient with high efficiency.