Molecular cloning, expression and vaccination of a novel gene Sj-MA of Schistosoma japonicum.

Yu-Xiao Chen, Xin-Yuan Yi, Xian-Fang Zeng, Shi-Shan Yuan, Shun-Ke Zhang, Larry McReynolds
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Abstract

In order to obtain new gene and to develop the new vaccine candidate of immunoprotection against Schistosoma japonicum (Sj), Sj adult worm cDNA library was screened with anti-sera to soluble male adult worm antigen and resulted in the discovery of a novel gene designated as Sj-MA. Sequence analysis showed that Sj-MA as a complete cDNA contains one open reading frame. It was deduced to contain 249 amino acid residues and encode a 28.8 kD soluble protein with plenty of phosphorylation sites, supposing its action in signal transduction. Furthermore, Sj-MA cDNA was cloned into a prokaryotic expression vector pGEX-5X to construct the recombinant plasmid which was transformed and highly expressed in E. coli as a 54.8 kD glutathione-S-transferase (GST) fusion protein. The fusion protein rSj-MA/GST could be recognized with both anti-male adult worm sera and anti-GST sera in Western blotting. Mice vaccinated with the fusion protein revealed significant worm reduction rate of 34.29% (P<0.001), compared with the control groups. Taken together, the novel gene Sj-MA can be expressed in E. coli as a fusion protein that can elicit immunity against Schistosoma japonicum, suggesting its potential as a new vaccine candidate.

日本血吸虫新基因Sj-MA的分子克隆、表达及疫苗接种。
为了获得日本血吸虫(Schistosoma japonicum, Sj)的新基因和开发新的免疫保护候选疫苗,用可溶性雄性成虫抗原的抗血清筛选了Sj成虫cDNA文库,发现了一个新的基因,命名为Sj- ma。序列分析表明,Sj-MA作为一个完整的cDNA包含一个开放阅读框。据推测,该蛋白含有249个氨基酸残基,编码一个28.8 kD的可溶性蛋白,并具有大量磷酸化位点,推测其在信号转导中起作用。将Sj-MA cDNA克隆到原核表达载体pGEX-5X中构建重组质粒,转化为54.8 kD谷胱甘肽- s转移酶(GST)融合蛋白,在大肠杆菌中高表达。融合蛋白rSj-MA/GST在抗雄性成虫血清和抗GST血清中均能被识别。接种融合蛋白的小鼠蠕虫减少率为34.29% (P
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