重组人甲状旁腺素(rhPTH(1-84))的原核表达及发酵条件初步优化

 甲状旁腺激素(Parathyroid Hormone,PTH)是治疗骨质疏松症的药物之一.将人工合成全长人PTH(hPTH(1-84))的核苷酸序列插入pThioHis A载体中,然后转化大肠杆菌(Escherichia coli.),在IPTG的诱导下,成功实现了rhPTH(1-84)的原核表达.通过发酵条件的优化,初步确定1:40接种LB+30%M9盐溶液的发酵培养基,37℃培养至OD600 nm=0.8时,加入终浓度为0.6 mmol/L的IPTG,诱导8 h的较优发酵程序.

Recombinant expression of human parathyroid hormone(1-84)and preliminary research on fermentation conditions

Parathyroid Hormone(PTH)is one of durgs which could treat osteoporosis.Full-length human PTH(hPTH(1-84))nucleotide sequence was artificially synthesized,and after digestion with Nde Ⅰ and Sal Ⅰ,the fragment was inserted into pThioHis A.Then the recombinant plasmid was transformed into E.Coli BL21(DE3).At last,the hPTH(1-84)was recombinantly expressed by inducing of the IPTG.After studying on fermentation conditions,the best expression could be obtained when BL21(DE3)cultures were diluted 1/40 into medium with ampicillin including LB and 30% M9 salt,and grown at 37℃ to an OD 600 nm of 0.6,finally added 0.6 mmol/L IPTG and incubated another 8 hours.