人精子蛋白SP17基因的克隆及其在大肠杆菌中的表达  

Cloning of human sperm protein(SP17) and expression in escherichia coli DH5α

作　　者：王敏珍[1] 刘志成[1] 高展[2] 郑文波[2] 罗建红[2] 

机构地区：[1]浙江大学医学院邵逸夫医院妇产科,浙江杭州310006 [2]浙江大学医学院医学分子生物学实验室,浙江杭州310006

出　　处：《中国病理生理杂志》2001年第10期1019-1021,共3页Chinese Journal of Pathophysiology

摘　　要：目的 :获得人精子蛋白SP17基因 (hSP17) ,构建重组表达载体并在大肠杆菌中表达。方法 :获取人睾丸组织 ,提取总RNA ,通过RT -PCR获得hSP17的全长cDNA ;将该cDNA克隆入TA载体 ,并亚克隆进融合蛋白型重组表达载体pGEX - 3b ;转化大肠杆菌DH5α并诱导表达之。结果 :获得了全长hSP17的cDNA ,构建了重组表达子hSP17/pGEX - 3b ,获得了预期大小的融合蛋白hSP17-GST的表达。结论 :通过构建重组表达质粒hSP17/pGEX并转化大肠杆菌 ,获得了融合蛋白hSP17-GST的高效表达。AIM: To obtain GST fusion protein of hSP17 gene and construct the recombinant plasmid for expression in E. coli. METHODS: Total fragment of hSP17 cDNA gene were amplified by RT-PCR, then subcoloned into pGEX-3b to generate recombinant hSP17/pGEX. Right orientation of insert are identified by restricted enzyme digestion. Transform the correct recombinant plasmid into the E. coli DH5a. The expression of fusion proteins hSP17-GST were induced by adding isopropylthiogalactoside (IPTG). RESULTS and CONCLUSION: The recombinant plasmid hSP17/pGEX-3b could express effectively in E.coli and a high level of fusion protein hsp17-GST with the predicted molecular weight was detected.

关 键 词：人类 精子 蛋白质类 SP17基因 克隆 大肠杆菌 基因表达 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学] Q786[医药卫生—基础医学]