Construction and Bioactivity of Recombinant Baculovirus with cry1Ac10 Gene of Bacillus thuringensis
Abstract: Using Bac-to-Bac system, the transfer vector pFCP with full-length cry1Ac10 gene of Bacillus thuringensis drived by the very late expressed gene ph promoter and intact polyhedra gene was constructed successfully. The recombinant virus vFcph , with complete polyhedra and cry1Ac10 gene, was obtained by transfection of the transfer vector pFCP DNA into insect Sf9 cells and cry1Ac10 protein can be expressed in insect cells. Meanwhile, a new shutter vector pHTC with cry1Ac10 gene was constructed. Three kinds of engineered bacteria produced by transformation of Bacillus thuringensis, Escherichia coli and Bacillus subtilis all expressed protoxin with the molecular weight of 133.3kDa. The expression of the protoxin in Bacillus thuringensis was the highest. Bioassay indicated that the expressing product can increase virulence and killing speed of baculovirus. The results indicated that it is feasible to enhance the effectiveness of baculovirus by inserting an insecticide crystal protein gene into baculovirus controlled by the promoter of very late expressed gene.