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| The Study On Construction, Immunogenicity and Protection Against Mycobacterium tuberculosis ofmpt64-recombinant BCG Vaccine |
| SUN Ying1,2, ZHANG Ling-xia3, WU Xue-qiong3, DONG En-jun3 |
1. Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 101300, China;
2. Beijing Technology Exchange & Promotion Center, Beijing 100080, China;
3. The 309th Hospital of PLA, Beijing 100091, China |
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Abstract It was aimed to construct mpt64-recombinant BCG vaccine to find an improved vaccine to replace BCG and to prevent TB effectively. The gene of MPT64 was amplified by PCR and recombined with shuttle plasmids pYUB295. The recombinant shuttle plasmid was identified by PCR, enzyme digestion and DNA sequencing and then transformed into BCG by electroporation. The antigen-specific antibody levels of mouse immunized with recombinant BCG were evaluated by ELISA and multiplication of mouse lymph-cell was detected by MTS. The protection against M. tuberculosis of recombinant BCG vaccines was tested by their prevention and treatment experiments. PCR, enzyme digestion, DNA sequencing and SDS-PAGE results showed: Thempt64-recombinant BCG was constructed successfully and can express extrinsic MPT64 gene. The immunity experiment showed:extrinsic gene can be expressed in BCG and can stimulate B cell to produce antibody,the antibody level reached the peak after 45 days; The lymph-cells of each group proliferated when stimulated by different antigen, all stimulation index reached 2. The stimulation index of each group had no obviously difference. The prevention experiment of recombinant BCG against M. tuberculosis was indicated: mpt64-recombinant BCG and BCG can extend mean time to death and reduce death rate in 2 month of those mouse, the protection effect of mpt64-recombinant BCG had no difference with BCG. It could be concluded that the mpt64-ecombinant BCG was constructed successfully. The protection effect of mpt64-recombinant BCG had no difference with BCG.
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Received: 16 November 2010
Published: 25 July 2011
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