人卵细胞ZP3蛋白在大肠杆菌中的表达与鉴定  

Expression and identification of human zona pellucida 3 protein in Escherichia coli

作　　者：李津[1] 谢飞[2] 涂向东[2] 郑德柱[2] 兰风华[2] 

机构地区：[1]第二军医大学福州总医院临床医学院,福建福州350025 [2]南京军区福州总医院全军医学检验中心,福建福州350025

出　　处：《东南国防医药》2006年第4期243-244,255,共3页Military Medical Journal of Southeast China

基　　金：全军医药"十五"基金项目(01MA032);福建省重点科技攻关项目(2002Y032)

摘　　要：目的构建人卵细胞ZP 3蛋白的原核重组载体并进行表达和鉴定。方法提取人卵巢中总RNA,逆转录合成cDNA,自行设计引物,PCR扩增ZP 3蛋白编码序列,PCR产物经B amH I/N heⅠ双酶切后,克隆至表达载体pET-H is中,在E.co li BL 21-DE 3中诱导ZP 3融合蛋白的表达。结果在异丙基-β-硫代半乳糖苷(IPTG)的诱导下,重组菌高效表达出一个相对分子质量约为50 000的产物。结论人卵细胞ZP 3蛋白的全长编码序列已被克隆至ZP 3蛋白融合表达载体pET-H is,并在E.co li BL 21-DE 3中获得高表达。Objective To construct a prokaryotic recombinant vector of human zona pellucida 3 protein and to detect its expression in E. eoli BL21-DE3. Methods Total RNA was extracted from human ovary tissue using RNeasy Mini Kit. After reverse transcription, the coding sequence of ZP3 was amplified with specific primers designed in our laboratory, and cloned into pET-His vector after the restrictive digestion with BamHI and Nhe Ⅰ . The his fusion protein was expressed after induction with IPTG. Results Sequencing and restrictive digestion of the recombinant plasmid revealed the existence of coding sequence of human ZP3 protein. A protein band of about 50 000 could be induced by IPTG in the recombinant plasmid. Conclusions The coding sequence of human ZP3 protein is introduced into the pET-his plasmid and a his fused protein can be induced in E. eoli BL21-DE3 at a high level.

关 键 词：人卵细胞ZP3蛋白 克隆 基因表达 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学]