弓形虫P30基因在昆虫杆状病毒载体体系中的初步表达

将弓形虫主要表面抗原（Ｐ３０）基因插入转移载体质粒ｐＳＸＩＶＶＩ＋Ｘ３中，然后将重组质粒ｐＳＸＩＶＶＩ＋Ｘ３－Ｐ３０ＤＮＡ与粉纹夜蛾核形多角体病毒ＴｎＮＰＶＤＮＡ共转染培养的草地夜蛾细胞（Ｓｆ），通过同源重组和空斑纯化，得到重组病毒ＴｎＮＰＶ－Ｐ３０．对重组病毒感染的Ｓｆ细胞及银纹夜蛾幼虫进行ＳＤＳ－ＰＡＧＥ和Ｗｅｓｔｅｒｎｂｌｏｔ分析。结果显示：Ｐ３０基因在Ｓｆ细胞及虫体中得到了表达，表达产物具有特异的免疫反应性。

The gene encoding the major surface antigen (P30) of Toxoplasma gondii was cloned into a transfer plasmid vector pSXIVVI +X3, then the recombinant plasmid pSXIVVI +X3-P30 DNA and the parent virus TnNPV DNA cotransfected the cultured Spodoptera frugiperda (Sf) cells. Through homo-recombination, the recombinant virus TnNPV-P30 was raised, and purified by plaque assay. SDS-PAGE and Western blot analysis of the proteins from Sf cells and Argyrogramma agnata larvae infected by TnNPV-P30, showed that P30 had been expressed in these Sf cells and larvae. The expression product appeared to react specifically with the rabbit serum against T. gondii .