The appearance of the first comericial plasmid pHT01 promotes the study for inducible over-expression of recombinant protein in Bacillus subtilis (B. subtilis). Most of researches published relating to this plasmid used B. subtilis 168 derivatives, such as 1012 strain, but not with WB800N - eight-fold protease-deficient strain. To explore the possibility to use plasmid pHT01 with B. subtilis WB800N for secretional over-expression of recombinant protein, the authors used secretional reporter protein, alpha-amylase encoded by amyQ from Bacillus amyloliquefasciens. Plasmid pHT43-amyQ was constructed from pHT01 and used to investigate the secretional expression of alpha-amylase in both B. subtilis 1012 and B. subtilis WB800N. The results showed that alpha-amylase could be produced efficiently in both strains, in which the amount of the reporter from WB800N was higher than that from 1012. This result suggests that plasmid pHT01 can be used with B. subtilis WB800N for secretional expression of recombinant protein.