摘要
人甲状旁腺激素(Humanparathyroidhormone,hPTH)是84个氨基酸组成的多肽激素。从人甲状旁腺肿瘤组织分离纯化总RNA,其mRNA逆转录得cDNA,以cDNA为模板,PCR扩增得hPTH基因。将该基因克隆到表达载体pBV220上,转化大肠杆菌DH5α细胞,获得了表达。细胞抽提液1000倍稀释后hPTH的放免活性26577ng/dl,为对照的480倍。表达产物粗分离后经ResourceS阳离子柱进行了分离。
Human parathyroid hormone(hPTH) is a peptide hormone consisting of 84 amino acids.Total RNAs were isolated from parathyroid adenoma tissue and mRNAs were reverse transcripted to obtain cDNAs.The hPTH gene was then amplified from cDNAs by PCR,inserted into expression vector pBV220 which contains a temperature inducible promoter and confirmed by DNA sequencing.The recombinant plasmid was used to transform E.coli DH5α.The radiommuno activity of hPTH in the crude extract of DH5α containing recombinant plasmid pGA555 was 265 77ng/dl,which is 480 times as that of control.After partial purification of the expression product,the supernatant was applied to Resource S column chromatography.The radiommuno active peak was collected and characterized by HPLC C 18 reverse phase column and MALDI mass spectrometric analysis.
出处
《生物工程学报》
CAS
CSCD
北大核心
1997年第3期264-268,共5页
Chinese Journal of Biotechnology
关键词
甲状旁腺激素
基因克隆
表达
纯化
基因工程
Human parathyroid hormone, E.coli ,gene cloning and expression,isolation and purification