Cloning of the Molecular Chaperone Gene p19-p29 from Bt and Construction of the Bt Expression Vector
苏云金芽孢杆菌分子伴侣串联基因p19-p29的克隆和表达载体的构建

The primer pair (ORF-5S,ORF-3N)designed according to the known sequence can amplify a DNA fragment, containing molecular chaperone genes p19 and p29, in size of 1.6kb from the cry2Aa operon or of 2.0kb from the cry2Ac operon. 150 Bacillus thuringiensis(Bt) strains were detected by PCR with the primer pair. With a blankness of the 2.0kb fragment, the expected 1.6kb fragment was acquired from 26 Bt strains. These results showed that the genotype of cry2Aa operon widely existed in Bt and that cry2Ac was rare. The 1.6kb PCR product from strain Y 2 was recovered. After several steps of sub-cloning, it was at last inserted into the shuttle vector pHT3101, resulting in an expression vector pHY 2P with multiple clone sites.