Cloning of prtR gene from bacillus subtilis

Abstract

In Bacillus subtilis, many genes responsible for hyperproduction of extracellular enzymes have been reported and new ones are expected to be exist. prt R gene, for example, encodes a 60-amino acid polypeptide and can enhance the transcription of neutral and alkaline proteases and levansucrases. To clone prt R gene or other genes which has pleiotropic effect, a system was developed in which those genes could be cloned by simple Cm-selection. But this system was turned out to be unreliable because small portion of cells showed greatly enhanced Cm-resistancy. Using $^{32}P$-labelled oligonucleotide as a probe, one colony showed positive signal in colony hybridization. The recombinant plasmid contained a 1.65 kb fragment. When subcloned into B. subtilis MI112 cells, moderate change of production of extracellular proteases was detected on skim-milk plate. Further characterization should be needed.