Expression of Stichopus japonicus Lysozyme Gene in Bacillus subtilis WB600

摘要

In this study, a genetic engineering bacteria Bacillus subtilis pHT43-SjLys/ WB600 was successfully constructed for the expression of the lysozyme gene from sea cucumber (Stichopus japonicus) by the method of recombinant DNA technique. The growth trend of engineering bacteria was consistent with the wild-type strain WB600, and the results demonstrated that insertion of foreign gene did not affect its physiological and biochemical metabolism. In the absence of selection pressure, the analysis of the stability revealed that there was no gene rearrangement and lost of the recombinant plasmid in the bacteria which showed that it has high genetic stability. The SDS-PAGE results demonstrated that pHT43-SjLys/WB600 successfully expressed soluble SjLys protein after incubated for 48 h induced by ITPG. The heterologous expression protein of pHT43-SjLys/WB600 displayed remarkable inhibitive effect on the growth of the Vibrio parahaemolyticus. To our knowledge, this is the first report about the SjLys gene authentic heterologous expression in B. subtilis. It should provide a robust secretion expression system for genetic engineering of B. subtilis and was thus proposed a potentially new way for producing recombinant SjLys protein.