日本血吸虫Rho GTPase DNA疫苗及亚单位疫苗的构建和分析

目的 构建日本血吸虫大陆株Sj-Rho GTPase-like真核及原核表达重组质粒，并进行鉴定分析。方法 用PCR法将Sj-Rho GTPase-like基因从已剪切阳性克隆中扩增出来，亚克隆至pcDNA3.1和pGEX-5X-3中，分别经PCR、双酶切、测序、SDS-PAGE和Western blot等方法鉴定。结果 PCR和测序均证明Sj-Rho GTPase-like基因疫苗构建成功，重组蛋白经SDS-PAGE电泳可观察到与预期分子量相应的条带，转印后可被水牛感染血清识别。结论 成功构建了Sj-Rho GTPase-like基因的两种重组载体，为保护性免疫研究打下基础。

Construction and analysis of Sj - Rho GTPase DNA vaccine and subunit vaccine against Schistosoma japonicum

Objective To construct a eukaryotic expression plasmid and a prokaryotic expression plasmid containing the gene encoding Sj-Rho GTPase-like of schistosoma japonicum. Metheds Sj-Rho GTPase-like gene obtained by PCR amplification from the positive clone had been excised and ligated with the plasmid pcDNA3.1 and pGEX-5X-3 separately. Recombinant plasmids were identified by PCR and restriction enzyme digestion and sequencing and SDS-PAGE and Western blot. Results The DNA vaccine was constructed successfully. It is identified by PCR and sequencing. The 51kD fusion protein was highly expressed in E.coli strain ER2566 induced by IPTG and recognized by infected serum of Bb. Conclusion The Sj-Rho GTPase-like gene was successfully cloned into the pcDNA3.1 and pGEX-5X-3 vector and could be used as DNA vaccine and recombinant protein vaccine respectively.