以pPIC9K为载体,构建抗HIV 1gp12 0单链抗体scFv12 0与葡萄球菌肠毒素A(StaphylococcalenterotoxinA ,SEA)融合基因表达质粒,线性化、电转化法整合入巴斯德毕赤酵母菌,经表型鉴定、PCR分析和G4 18筛选得到Muts型多拷贝整合菌,甲醇诱导培养可分泌表达5 7kD的预期大小蛋白—重组导向毒素SL12 0 ,表达量达5 0 1mg L。通过单链抗体亲和力测定,表明蛋白SEA和scFv12 0的构象有微弱的相互影响,但此重组导向毒素仍可高效介导CTLs杀伤HIV 1靶细胞。 The pPIC9K vector was used to construct a fusion gene expression plasmid for scFv12 0 against Staphylococcalenterotoxin A (SEA), which was integrated into the Pichia pastoris by the linearization and electrotransformation method. Identification, PCR analysis and G4 18 screening Muts type multi-copy integrated bacteria, methanol induced culture can secrete 5 7kD expected size protein - recombinant toxin SL12 0, the expression level of 501 mg. The single-chain antibody affinity assay showed a weak interaction between the protein SEA and scFv12 0 conformations, but this recombinant toxin still efficiently mediated CTLs to kill HIV 1 target cells.