弓形虫主要表面抗原基因的克隆

Amplification、clone、sequence and expression of the gene encoding the major surface antigen of Toxoplasma gondii isolated from China

根据弓形虫主要表面抗原（Ｐ３０）的基因序列合成了一对引物，应用聚合酶链反应技术从弓形虫中国分离株（ｚｓｌ）中，扩增了Ｐ３０的编码基因。序列测定显示该基因序列与国际标准弓形虫株（ＲＨ）的Ｐ３０基因序列几乎完全一致。扩增的Ｐ３０基因被克隆到质粒ｐＢＶ２２０中。ＳＤＳ－ＰＡＧＥ和免疫印迹分析显示，Ｐ３０基因在大肠杆菌中不仅有表达，且表达产物具有特异的免疫反应性。

According to the published gene sequence of the major surface antigen(P30), a pair of primers weredesigned and synthesizedo. Using the PCR , the coding sequences of P30 gene were amplified from a chi-nese strain of Toxoplasma gondii. The amplified gene fragments and plasmid pBV220 were digested withEcoRI and BamHI and then ligated. The inserted gene fragment was sequenced by the chain terminationmethod, the reading reveal that nucleotide sequence determinded here was the same as the P30 sequence ofRH stain published by Burg(1988) , except one base was exchanged. The recombinant plasmid containingP30 gene was transformed to E coli DH5a. After temperature inducing culture,the total cellular proteinswere analysed by SDS- PAGE and western blot. The results show the P30 gene cloned into the plasmid notonly expressed in E coli, but also the expression product appear a specific immunogenicity.