人脂联素基因的克隆及在大肠杆菌中的表达  

Study on the Clone of Human Adiponectin Gene and Its Expression in Escherichia Coli

作　　者：刘德敏[1] 杨莉丽[1] 孙颖[1] 张捷[1] 

机构地区：[1]天津医科大学代谢病医院,300070

出　　处：《天津医药》2006年第3期148-150,共3页Tianjin Medical Journal

基　　金：天津市自然科学基金资助项目(项目编号:20030325)

摘　　要：目的:克隆人脂联素(adiponectin)基因,并在大肠杆菌宿主系统中高效表达出adiponectin蛋白进行鉴定。方法:从人皮下脂肪组织提取总RNA,确认其正确性后经RT-PCR扩增得到全长cDNA片段,此片段经纯化回收后克隆至pMD18-T载体,经行DNA序列分析,进一步克隆构建到表达载体至pET-DEST42中,用IPTG在大肠杆菌中诱导表达。结果:含重组adiponectin质粒的大肠杆菌经0.4mmol/L的IPTG诱导6h后有高表达。结论:adiponectin基因的克隆构建和在大肠杆菌中的表达获得成功。Objective： To clone human adiponectin gene and identify its expression in E.coli. Methods： Total RNA was extracted from human subcutaneous adipose tissue and a full length cDNA was obtained by RT-PCR. The product was ligated to pMD-18T vector, and was sequenced and constructed to the expression vector in pET-DEST 42, and the expression was induced by IPTG in E.coli. Results： A high level expression was obtained by 04mmol/L IPTG inducement in recombined adiponectin E. coil Conclusion： Human adiponectin gene in E.coli is successfully cloned and expressed.

关 键 词：胞间信号肽类和蛋白质类 克隆 分子 基因表达 大肠杆菌 

分 类 号：R589.2[医药卫生—内分泌] R378.21[医药卫生—内科学]