草原兔尾鼠GST-LZP3融合蛋白的原核表达及其抗体制备

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 ,为采用免疫不育技术进行草原兔尾鼠生育控制的研究提供了重要的制剂

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

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.