论文部分内容阅读
为发现新基因 ,寻找日本血吸虫病新疫苗候选分子 ,采用Sj雄虫免疫血清筛选Sj成虫cDNA文库。经测序发现新基因Sj MA含有一个完整的阅读框 ,推测其由 2 4 9个氨基酸组成 ,编码分子量为 2 8.8kD的可溶性蛋白质 ,并带有多个能被磷酸化激活的位点 ,提示其可能为一重要的信息传递分子。将Sj MA的cDNA亚克隆至原核表达载体pGEX 5X ,获得Sj MA原核表达的重组体rSj MA/GST ,并在E .coli中高效表达为谷胱甘肽S 转移酶 (GST)融合蛋白 ,分子量为 5 4 .8kD ,Western印迹显示融合蛋白质能被抗雄虫和抗GST血清识别。融合蛋白质免疫小鼠可诱导 34.2 9%的减虫率 ,与对照组有显著性差异 (P <0 .0 0 1 )。表明新基因Sj MA表达的蛋白质能诱导小鼠的抗日本血吸虫的保护性免疫 ,提示其作为日本血吸虫疫苗候选分子的潜在价值
In order to find new genes, we searched for new vaccine candidate genes of schistosomiasis japonica, and used Sj male immune serum to screen the Sj adult cDNA library. Sequencing revealed that the novel gene Sj MA contains a complete reading frame, which is presumed to be composed of 299 amino acids and encodes a soluble protein of 28.8 kD in size with multiple sites that can be activated by phosphorylation, suggesting that May be an important information transmission molecule. The cDNA of Sj MA was subcloned into the prokaryotic expression vector pGEX 5X to obtain the recombinant rSj MA / GST prokaryotic expression vector of Sj MA, which was highly expressed in E.coli as a glutathione S transferase (GST) fusion protein. The molecular weight At 54.8 kD, Western blotting revealed that the fusion protein was recognized by both anti-male and anti-GST sera. The fusion protein immunized mice induced a 34.29% reduction in worm reduction rate, which was significantly different from the control group (P <0.01). The results showed that the protein expressed by the novel gene Sj MA could induce the protective immunity of mice against Schistosoma japonicum, indicating its potential value as a candidate molecule of Schistosoma japonicum vaccine