【摘要】 根据曼氏血吸虫肌动蛋白基因SmAct2设计一对引物 ,以日本血吸虫中国大陆株成虫mRNA为模板 ,用RT PCR法扩增出一大小为 1131bp的基因片段。测序后分析序列推断该片段为编码肌动蛋白基因的完整阅读框基因 ,与SmAct2碱基同源性为 92 %。将其克隆到表达载体 pET2 8a(+)中 ,在大肠杆菌BL2 1中获得了较高量的表达 ,融合表达产物分子量约为 47kD。利用日本血吸虫成虫粗抗原免疫大白兔所得抗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 ,说明所克隆的基因为编码日本血吸虫中国大陆株肌动蛋白基因。更多还原

【Abstract】 A 1 131 bp cDNA fragment was amplified by RT-PCR from adult Schistosoma japon icum (Chinese strain) mRNA with primers designed according to published SmAct2 encoding Schistosoma mansoni actin. Sequence analysis indicated t hat this fragment, with 92% homology to SmAct2, was a complete open reading fragment (OR F) of actin gene of Schistosoma japonicum (Chinese strain). This gene was cl oned into the expression vector pET28a(+) and subsequently expressed in Escerichi a coli. SDS-PAGE revealed that the molecular weight of this expressed product was 47 kD .Western blotting showed that the recombinant protein had good reacti vity with the rabbit serum immunized with Sj worm antigen, indicating that t his gene encode actin of Schistosoma japonicum (Chinese strain).更多还原