Abstract: A versatile Escherichia coli-Bacillus subtilis shuttle vector pY980coli was constructed by genetic recombination. Resistance gene and replicon of pUC18, a cloning vector for Escherichia coli, were integtated into the Bacillus subtilis expression plasmid pWB980. Protease gene hspa from a marine bacterium was then inserted under P43 promoter of pY980coli, and the recombinant vector was transformed into Bacillus subtilis WB600. Protease activity was detected in the fermentation broth.