摘要
化学合成人甲状旁腺激素 (hPTH)全长基因 ,克隆到大肠杆菌表达载体pBV2 2 0和pET2 2b中 ,获得了高表达。经破菌、阳离子交换层析、反相层析纯化获得了纯度大于 95 %的纯品。N端测序、质谱分析结果表明重组hPTH结果完整 ,N端无Met或fMet。生物活性试验证明重组hPTH具有激活腺苷酸环化酶、增加骨质量和骨密度等作用。
Human parathyroid hormone(hPTH) was highly expressed in Escherichia coli by inserted the synthesized whole hPTH cDNA into the vectors pBV220 and pET22b.After expression and disruption,the purified product was acquired through cation exchange chromatography and reverse phase chromatography.From the results of N terminal sequencing and MALDI TOF MS analysis the recombiant prtein was indentified as intact hPTH.In in vitro Bioassays the recombinant hPTH stimulated adenylate cyclase as the standard did. In ovariectomized rats the recombinant hPTH markedly increased the femoral bone mass and bone mineral density.
出处
《生物工程学报》
CAS
CSCD
北大核心
2003年第1期102-106,共5页
Chinese Journal of Biotechnology
基金
军事医学科学院开发基金
国家 863项目资助 (No .2 0 0 1AA2 15 45 1 3 )~~
关键词
人甲状旁腺激素
大肠杆菌
表达
鉴定
human parathyroid hormone, Escherichia coli , gene expression
