Construction of Vector for Expression of hsp65 Gene Driven by Maize Endosperm-specific Promoter

The DNA of Hsp65 amplied from pEGHLE by polymerase chain reaction (PCR) were cloned into the vector pCRG. The combine fragment of promoter globulin and the target gene Hsp65 which got from doubled enzymes digestion of the recombined plasmid pCRGHsp65 was inserted into the plant express vector pCAMBIA1300 which contain the bar gene for herbicide resistance. The recombinant plasmid was analyzed by restriction enzyme digestion and the inserted target genes in the pC1300GHsp65 were verified by nucleotide sequencing. Then was transformed pC1300GHsp65 vector into Agrobacterium Tumerfaciens LBA4404 by electroporation. The results indicated that the binary expression plasmid, which could express Hsp65, was correctly constructed after detected by restriction enzyme. The result showed that the recombinant vector containing Mycobacterium tuberculosis Hsp65 was constructed successfully and transformed into LBA4404, and laid a foundation for further study on its immunity effectiveness against MTB.