Construction of Vector for Expression of Esat-6 Gene Driven by a Maize Endosperm-specific Promoter

To construct the plant expression plasmid containing Mycobacterium tuberculosis Esat-6 genes,and transform the recombined vector into Agrobacterium tumerfaciens LBA4404.The DNA of Esat-6 amplied from pEGHLE by polymerase chain reaction(PCR) were cloned into the vector pCRG2.1,The combine fragment of promoter globulin and the target gene Esat-6 which get from doubled enzymes digestion of the recombined plasmid pCRGEsat-6 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 pC1300GEsat-6 were verified by nucleotide sequencing.Then transformed pC1300GEsat-6 into Agrobacterium tumerfaciens LBA4404 by electroporation.The binary expression plasmid,which could express ESAT-6,was correctly constructed by digestion and sequencing.Digested plasmid which was extracted from Agrobacterium tumerfaciens,fragments were also correct.The recombinant vector containing Mycobacterium tuberculosis Esat-6 is constructed successfully and transformed into LBA4404,and lays a foundation for further study on its immunity effectiveness against MTB.