RECOMBINANT SPORE WITH BOMBYXMORI ALCOHOL DEHYDROGENASE DISPLAYED ON ITS SURFACE AND PREPARING METHOD THEREOF

摘要

Bombyxmorialcoholdehydrogenase is displayed as exogenous protein on the Bacillus subtilis surface. Recombinant integrative plasmid for fusion expression of Bombyxmorialcohol dehydrogenase BmADH is constructed by using cotC, the gene of Bacillus subtilis spore coat protein, as molecule carrier, and being the latter recombined with Bombyxmori alcohol dehydrogenase gene BmADH. Bacillus subtilis is transformed with the plasmid, and the exogenous gene is integrated into the amylase gene amyE at the specific site onchromosome by homologous recombination, thus the amylase gene is destroyed and amylase activity of the recombinant strain is absent, therefore the recombinant Bacillus subtilis strain comprising fusion expression of CotC-BamADH is selected. The spore with Bombyxmori alcohol dehydrogenase fusion protein displayed on its surface is obtained by inducible expressionof recombinant strain. The alcohol dehydrogenase unstable at certain temperature and pH originallyis constructed into alcohol dehydrogenase fusion protein with relatively higher stability at different environment conditions, and the recombinant spore can be orally administered to animals directly.