华支睾吸虫成虫RPEF基因产物及其抗RPEF多克隆抗体的制备

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pGEx4T-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。

The preparation of RPEF gene products of Clonorchis sinensis and the anti-RPEF polyclonal antibodies

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.