猪卵透明带-3β蛋白核心片断在E.coli中的表达  

Expression of Core Domain of Porcine Zona Pellucida Protein 3β in E.coli

作　　者：谢秋玲[1] 陈小佳[1] 朱伟杰[2] 张玲[3] 徐万祥 洪岸[1] 李菁[5] 高丝红[1] 

机构地区：[1]暨南大学生物工程研究所,广州510632 [2]暨南大学生殖免疫研究中心,广州510632 [3]广东省生物工程药物重点实验室,广州510632 [4]上海市计划生育科研所,上海200032 [5]暨南大学医学院病理生理教研室,广州510632

出　　处：《生殖与避孕》2005年第7期387-390,共4页Reproduction and Contraception

基　　金：广东省科技计划项目(2001C12001)

摘　　要：目的:探讨通过基因工程的方法在大肠杆菌中表达猪卵透明带蛋白pZP3b的核心片断。方法:采用PCR方法扩增原cDNA中编码第44-306个氨基酸的pZP3β核心片断的DNA序列,克隆入pET-3c载体进而在大肠杆菌E.coliBL21(DE3)pLysS中表达。结果:表达的pZP3β核心蛋白占菌体总蛋白的15%,并经Western-blot鉴定为目的蛋白。结论:pZP3β核心片断在大肠杆菌中获得较高的表达,有利于产物的纯化,为后续阐明pZP3β核心区域的性质和相关抗生育研究打下了基础。Objective： To obtain the recombinant core domain of procine zona pellucida protein 3β （cZP3β） for further research in its function. Methods： By using PCR, the nucleotides sequence region from 44 to 306 codons of pZP3β entire coding cDNA fragment was obtained. Such sequence was cloned into pET-3c vector, then was transformed into the E.coli BL21 （DE3）pLysS and induced by IPTG to express the exogenous protein. Results： The recombinant cZP3β was expressed in E.coli up to 15% of total cellular protein, and was made sure by Western-blot analysis. Conclusion： High expression of core domain of pZP3β could be obtained in E.coli, which would benefit further investigation for its immunogenicity.

关 键 词：猪卵透明带-3β 核心片断 大肠杆菌 表达 

分 类 号：Q78[生物学—分子生物学]