Abstract: To construct the plant expression plasmid containing Mycobacterium tuberculosis Hsp65 and Esat26 genes, and transform the recombined vector into Agrobacterium tumerfaciens LBA4404. The fusion DNA fragment of Hsp65 and Esat26 were amplied from pEGHLE by polymerase chain reaction(PCR)were cloned into the vector pCRG. The combine fragment of promoter globulin21 and the target gene HLE which get from doubled enzymes digestion of the recombined plasmid pCRGHLE was inserted into the plant expression vector pCAMBIA1300 which contain the gene bar for herbicide resistance. The recombinant plasmid was analyzed by restriction enzyme digestion and the inserted target genes in the pC₁300GHLE were verified by nucleotide sequencing. Then transformed pC₁300GHLE vector into Agrobacterium tumerfa2 ciens LBA4404 by electroporation. The binary expression plasmid,which could express HSP65 and ESAT26,was correctly constructed. The recombinant vector containing Mycobacterium tuberculosis Hsp65 and Esat26 genes is constructed suc2 cessfully and transformed into LBA4404,and lays a foundation for further study on its immunity effectiveness against MTB.

Key words: Mycobacterium tuberculosis, Hsp65, Esat26, globulin21

CLC Number: 

• Q786