草原兔尾鼠GST-LZP3融合蛋白的原核表达及其抗体制备

Expression of Lagurus ZP3 fusion protein in prokaryotic cells and preparation of its antiserum

目的 :在原核中表达并纯化草原兔尾鼠卵透明带 3 (zonapellu cida3 ,ZP3 ) ,并以其作为抗原 ,制备抗ZP3的抗血清。方法 :将LZP3基因的核心片段克隆到原核表达载体pGEX 4T 1中。经酶切和序列分析后 ,用重组质粒转化大肠杆菌BL2 1(DE3 ) ,并经丙基β D 硫代半乳糖苷 (IPTG)诱导产生GST LZP3融合蛋白。以纯化的融合蛋白免疫新西兰兔制备抗血清。抗血清的效价及特异性采用ELISA和Westernblot检测。结果 :成功地构建GST LZP3融合蛋白表达载体 ,并在大肠杆菌中高效表达特异性的GST LZP3融合蛋白。以该融合蛋白免疫兔子获得抗GST LZP3融合蛋白的高效价抗血清。结论 :获得原核表达的GST LZP3融合蛋白并制备了兔抗LZP3的抗血清 。

AIM: To express and purify Lugurus zone pellucida 3 (LZP3) in prokaryoti c cells and to prepare the LZP3-specific rabbit antiserum. METHODS: The core fragment of LZP3 gene was cloned into plasmid pGEX-4T -1 containing glutathione s-transferase(GST) fusion protein gene. Following re striction enzyme digestion analysis and sequencing, pGEX-4T-LZP3 w as transformed into E.coli BL21(DE3). GST-LZP3 fusion protein was expressed under IPTG induction and further purified with Glutathione Sepharose 4B. Then the purified GST-LZP3 fusion protein was used to immunize New Zealand rabbits. LZP3-specific rabbit antiserum was identified by ELISA and Western blot. RESULTS: GST-LZP3 fusion protein was overexpressed and its LZP3-spe cific antiserum was obtained. CONCLUSION: The successful express ion of GST-LZP3 fusion protein in E.coli and the preparation of LZP3-spec ific rabbit antiserum will be valuable for the study on birth control of Lagu rus.