| 华支睾吸虫成虫RPEF基因产物及其抗RPEF多克隆抗体的制备 |
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| 引用本文： | 张咏莉,吴德,胡旭初,郑焕钦,徐劲,余新炳. 华支睾吸虫成虫RPEF基因产物及其抗RPEF多克隆抗体的制备[J]. 国际免疫学杂志, 2002, 32(1): 251-254. DOI: 10.3760/cma.j.issn.1673-4394.2009.04.001 |
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| 作者姓名： | 张咏莉 吴德 胡旭初 郑焕钦 徐劲 余新炳 |
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| 作者单位： | 广东药学院基础学院生物教研室,广州,510224;广东省疾病控制中心,510300;中山大学基础医学院病原生物学部寄生虫教研室,510089; |
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| 基金项目： | 广东省自然科学基金团队项目"211"重点学科建设基金广东药学院2005年度科研项目 |
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| 摘 要： | 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.
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| 关 键 词： | 华支睾吸虫 RNA聚合酶Ⅱ延长因子 基因产物 抗血清 多克隆抗体 RNA polymerase Ⅱ elongation factor (RPEF) |
The preparation of RPEF gene products of Clonorchis sinensis and the anti-RPEF polyclonal antibodies |
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| Abstract: | 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. |
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| Keywords: | Clonorchis sinensisGene productsAntiseraPolyclonal antibody |
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