Fang Hong-Qing, Dai Hong-Mei, Li Yan-Ying, Zhao Hong-Liang, Deng Bing-Bing, Xue Chong, Liu Zhi-Min, Zhu Hou-Chu, Ma Qing-Jun, Chen Hui-Peng
Beijing Institute of Biotechnology, Beijing 100071, China.
Sheng Wu Gong Cheng Xue Bao. 2003 Jan;19(1):102-6.
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.
通过将合成的完整人甲状旁腺激素（hPTH）cDNA插入载体pBV220和pET22b中，使人甲状旁腺激素在大肠杆菌中高效表达。表达并裂解后，通过阳离子交换色谱和反相色谱获得纯化产物。通过N端测序和基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）分析结果，鉴定重组蛋白为完整的hPTH。在体外生物测定中，重组hPTH与标准品一样刺激腺苷酸环化酶。在去卵巢大鼠中，重组hPTH显著增加了股骨骨量和骨矿物质密度。
