日本血吸虫中国大陆株抱雌沟蛋白编码基因的克隆和表达

根据曼氏血吸虫抱雌沟蛋白SmGcp序列和日本血吸虫编码抱雌沟蛋白保守区的基因片段jGcp1分别设计三对引物，以日本血吸虫中国大陆株成虫mRNA为模板 ，用RT－PCR法扩增出大小为1949bp的基因片段。经序列分析推断该基因片段含编码日本血吸虫抱雌沟蛋白基因的阅读框，与SmGcp碱基一致性为85％，其理论推测氨基酸组成与曼氏血吸虫抱雌沟蛋白的一致性为83．7％。将上述扩增的基因片段克隆到表达载体pET28c( )中，在大肠杆菌BL21中获得表达，融合表达产物分子量约为80kD。利用日本血吸虫成虫抗原免疫血清对该表达产物进行Western印迹检测，在预测位置出现了明显的识别条带，说明该编码日本血吸虫中国大陆株抱雌沟蛋白基因的表达产物具有抗原性。

Cloning and Expression of Gynecophoral Canal Protein Gene of Schistosoma japonicum (Chinese strain)

A 1949 bp cDNA fragment was amplified by RT-PCR from adult Schistosoma japonicum (Chinese strain) mRNA with 3 pair of primers that were designed according to published SmGCP gene encoding gynecophoral canal protein of Schistosoma mansoni and SjGCP1 gene encoding the conservative region of gynecophoral canal protein of Schistosoma japonicum. Sequence analysis indicated that this fragment, named SjGCP, with 85% identity to SmGCP, contained a complete open reading frame (ORF) of gynecophoral canal protein gene of Schistosoma japonicum (Chinese strain). The amino acid sequence shared 83.7% identity with gynecophoral canal protein of Schistosoma mansoni. This fragment was cloned into the expression vector pET28c( ) and subsequently expressed in Escherichia coli. SDS-PAGE revealed that the molecular weight of this expressed product was 80 kD. Western blotting showed that the recombinant protein reacted well with the rabbit serum immunized with Sj worm antigen, indicating that this expressed product had good antigenicity.