关键词: 弓形虫, 多表位基因, 番茄果实特异性启动子, 植物表达载体

Abstract: 　Objective To construct the plant expression vectors containing the multiepitope gene of Tazoplasma gondii (TGMG). Methods ① TGMG was subcloned into pBAC55 vector to construct the intermediate plasmid pB35MG. The E35S/TGMG/NOS3' fragment was cleaved from pB35MG and ligated into the plant binary vector pCAMBIA2300 to construct the plant expression vector pC35MG. ② Tomato fruit-specific E81. 1 promoter was introduced to pB35MG to construct pB35E!MG vector. The E35SE81.1/TGMG/NOS3' fragment was subcloned into pCAMBIA2300 to construct the plant expression vector pCSSElMG. ③ Tomato fruit-specific E82.2 promoter was inserted to pB35MG to construct pBE2MG vector. The E82.2/TGMG/NOS3' fragment was subcloned into pCAMBIA2300 to construct the plant expression vector pCEZMG. The insert gene TGMG in the vectors pB35hKjs pCSSElMG and pCE2MG were confirmed by sequencing. ④ pC35MG, pCSSElMG and pCE2MG were introduced into Agrobacterium tumefaciens strain LBA4404 competent cell. Results Digestion with restriction enzymes proved that all recombinant vectors had the inserts with expected length of the target fragments. And the sequencing results were confirmed correct. Conclusion The TGMG intermediate vectors pB35MG, pBSSElMG and pBE2MG and the plant expression vectors pC35MG, pCSSElMG and pCE2MG were constructed successfully, and the three plant expression vectors were introduced into Agrribacterium tumefaciens.

Key words: Toxoplasma gondii, multiepitope gene, tomato fruit-specific promoter, plant expression vector