CAJ | 학술논문

目的体外制备华支睾吸虫成虫RNA聚合酶Ⅱ延长因子(RPEF)基因产物及抗RPEF多克隆抗体.方法亲和纯化试剂盒纯化表达蛋白pGEX-4T-1-RPEF,凝血酶酶切表达蛋白制备重组蛋白RPEF,将重组蛋白RPEF免疫小鼠,制备多克隆抗血清,检测抗体滴度和抗血清特异性.结果体外制备的RPEF蛋白经SDS电泳呈单一条带;酶联免疫吸附结果显示,免疫过程抗体滴度呈连续上升趋势;免疫印迹结果显示,小鼠抗血清具有抗RPEF特异性.结论获得较专一的华支睾吸虫RPEF蛋白和RPEF抗血清.
목적 체외제비화지고흡충성충RNA취합매Ⅱ연장인자(RPEF)기인산물급항RPEF다극륭항체.방법 친화순화시제합순화표체단백pGEX-4T-1-RPEF,응혈매매절표체단백제비중조단백RPEF,장중조단백RPEF면역소서,제비다극륭항혈청,검측항체적도화항혈청특이성.결과 체외제비적RPEF단백경SDS전영정단일조대;매련면역흡부결과현시,면역과정항체적도정련속상승추세;면역인적결과현시,소서항혈청구유항RPEF특이성.결론 획득교전일적화지고흡충RPEF단백화RPEF항혈청.
Objective To prepare RPEF Gene products of Clonorchis sinensis and the anti-RPEF polyclonal antibodies. Methods The expressed protein pGEX-4T-1-RPEF was purified by GST Bind Purification kit and was digested with thrombin. The experimental mice were inoculated with recombinant RPEF protein. The poly-clonal antisera were obtained from the immuned mice. The antisera collected were used to detect the titres of IgG antibodies and to identify anti-RPEF specificity. Results SDS-PAGE showed single band of the purified RPEF protein. ELISA analysis showed that IgG titer in the immuned mice was increased. The results of West-ern blot suggested that the mouse antir RPEF antisera were highly specific. Conclusion The RPEF gene products of Clonorchis sinensis and the anti-RPEF polyclonal antibodies were successfully prepared.