9316114 Jonas This project will result in the identification of genes transcribed by alternate sigma factors in the Gram positive prokaryote, Bacillus subtilis. Genes having promoters recognized by a particular sigma factor will be identified using a transposon engineered to contain a reporter gene (Tn917lac), which should generate random insertional mutations (and transcriptional fusions) in the chromosome of B. subtilis. These mutations will be screened in strains of B. subtilis containing inducible copies (under control of the spac promoter) of the structural genes for the active forms of B. or E. genes bearing insertion that are induced by IPTG in such a system will be mapped, cloned, and sequenced. The promoter regions of these genes will be analyzed, providing information concerning regulatory sequences used by alternative sigma factors. The functions of these genes may be revealed by the phenotype of the insertional mutants, or by homology with the sequences of genes with known functions. We will attempt to determine the role the alternative sigma factor plays in vivo by recognition of the function of the genes in the regulon. %%% The knowledge gained concerning genetic regulation may prove useful to the fermentation industry, which could benefit from using alternate sigma factors in over producing gene product in bacteria. ***
