中文摘要：

目的构建、表达并纯化polycomblike（Pcl）6xHis融合蛋白，制备该融合蛋白的大鼠多克隆抗体，纯化并鉴定该多克隆抗体。方法通过PCR反应从野生型果蝇形W^1118成虫cDNA中扩增Pcl基因部分片段，构建pRSETA-Pcl重组质粒。重组质粒转化大肠杆菌，IPTG诱导表达，Ni—NTA superflow层析柱纯化该融合蛋白，Western—blotting分析检测。用纯化的融合蛋白免疫SD大鼠，制备抗Pcl的多克隆抗体，溴化氰活化琼脂糖凝胶4B偶联法纯化多克隆抗体，Western-blotting检测该抗体的效率及特异性。结果成功构建原核表达质粒pRSETA-Pcl，大量表达并纯化Pcl融合蛋白。免疫大鼠获得多克隆抗体并纯化，Western-blotting及免疫组织化学染色结果表明该抗体可以检测Pcl全长蛋白及内源性蛋白。结论本研究获得了纯化的Pcl融合蛋白，成功制备并纯化了特异性较高的抗Pcl多克隆抗体，为进一步研究Pcl基因的功能奠定了基础。

英文摘要：

Polycomblike （Pcl） as a member of polycomb group, plays an important role in regulating the development of hemocytes, phagocytosis and antimicrobial peptides expression in the Drosophila. In this study, we aimed to prepare rat polyclonal antibodies against Pcl. A fraction of Pcl gene was amplified by PCR, and inserted into pRSETA vector. Then the pRSETA-Pcl plasmid was transformed into E. coli JM109 （DE3）, and the fusion protein was expressed under induction of IPTG. Then, the 6×His-Pcl protein were purified by Ni-NTA Superflow chromatography and used to immunize SD rats for generating polyclonal antibodies. The generated antibodies were purified with CNBr Activated Sepharose 4B, and then analyzed by Western blotting and immunofluorescence. The results indicated that the anti-Pcl antibodies are with high sensitivity and specificity, also can detect both Pcl full length and endogenous proteins, which would benefit the further function study of Pcl gene.