日本血吸虫卵壳蛋白编码基因的扩增和克隆

Amplification and Cloning of Eggshell Protein Gene of Schistosoma Japonicum

目的 克隆日本血吸虫卵壳蛋白基因 ,以研究其在诊断和作为候选疫苗分子的作用。方法 设计合成引物 ,以日本血吸虫雌性成虫cDNA第一链为模板 ,用PCR法扩增日本血吸虫卵壳蛋白 (SchistosomajaponicumeggshellproteinSjEP)基因编码序列 ,并克隆入pGEM T质粒载体。结果 PCR法扩增出大小为 62 4bp的单一条带 ,将其克隆入载体获重组质粒 pGEM T SjEP ,经双酶切和以质粒为模板进行PCR扩增 ,均可获得一条与PCR产物一致的DNA片段。结论 日本血吸虫卵壳蛋白的编码基因重组质粒的成功构建 ,为进一步研究提供了条件。

Objective\ To amplify eggshell protein(EP) gene of Schistosoma japonicum and to construct recombinant plasmids including SjEP gene for future study. Methods\ Primers were designed and synthesized. The DNA encoding eggshell protein of Schistosoma. japonicum was amplified by using polymerase chain reaction with female SjcDNA as template. The product from PCR was cloned into pGEM T vector. Thd PCR. Results\ For PCR, a specific sequence of 624bp was amplified. The same band was obtained through double restriction of recombinant plasmids and PCR with the recombinant plasmids used as template. Conclusion\ The present work may provide a basis for the further study of Schistosoma japonicum eggshell protein expression and its potential value for immunological diagnosis and prevention.