Construction and Identification of the Expression Plasmid Containing Genes Encoding the Large Envelope Protein L of HBV and Esat-6 Gene of Mycobacterium tuberculosis

To construction and identification of the expression plasmid containing genes encoding East6 gene of Mycobacterium tuberculosis and the large envelope protein L of HBV,and transform the recombinant vector into Agrobacterium tumefaciens LBA4404.The L and East-6 gene were amplied from pPIC9K-L and Mycobacterium tuberculosis genome by PCR,then the fusion DNA fragment of L and East6 were amplied by Splicing by Overlap Extension were cloned into the vector pEGG.The combine fragment of promoter globulin-1 and the target gene L-East6 which get from doubled enzymes digestion of the recombined plasmid pEG-G-L-Esat6 was inserted into the plant expression vector pCAMBIA1300 which contain the gene bar for herbicide resistance.Then transformed recombinant plasimid pCAM-G-L-Esat6 into Agrobacterium tumerfaciens LBA4404.We have successfully constructed eukaryotic expression recombinantplasmid pCAMG-L-Esat6 and it is showed that the cloned sequence of L and Esat6 is correct by sequencing.We have successfully constructed expression plasmid containing genes encoding L protein of HBV and Esat6 gene and transformed into LBA4404,and lays a foundation for construction combined gene vaccine against HBV and MTB.