基因工程猪生长激素复性技术的研究  

Renaturation of Recombinant-DNA-Derived Porcine Somatotropin

作　　者：于瑞嵩[1] 李震[1] 张平[1] 王英[1] 刘惠莉[1] 张德福[1] 

机构地区：[1]上海市农业科学院畜牧兽医研究所

出　　处：《中国生物工程杂志》2003年第6期64-67,共4页China Biotechnology

基　　金：上海市科技兴农重点攻关项目 (农科攻字 97第 5 -0 1号 );上海市现代生物与新药产业发展基金项目 (9843 190 0 2 )

摘　　要：对基因工程猪生长激素 (r pGH)体外复性条件进行研究 ,提取包涵体用 6mol L盐酸胍溶解 ,空气氧化 80h后转入复性缓冲液中开始重组蛋白复性 ,结果表明 ,采用透析复性法———复性缓冲液Ⅲ ( 0 2 5 %Na2 CO3 ,0 2 %乳糖 ,0 2 %甘露醇 ) ,pH8 5 ,透析 5～ 6次 ,可较好恢复r pGH生物活性。β 巯基乙醇、谷光苷肽等添加剂对r pGH的体外复性影响不显著。复性后蛋白浓缩液进行去垂体大鼠试验 ,大鼠增重明显 ,表明得到了正确折叠的较高生物活性的r pGH。The conditions for the in vitro renaturation of r pGH have been studied.The inclusion bodies extracted from host cells were dissolved using 6mol/L guanidine/HCl and air oxidation was carried out in the presence of guanidine/HCl.The guanidine/HC protein mixture was then diluted by renaturation solution.Guanidine/HC were removed by dialysis against renaturation solution Ⅲ(0.25% Na\-2CO\-3,0.2% lactose,0.2% mannitol,pH8.5) for 5 to 6 times and correctly refolded,oxidized r pGH were obtained.Injection experiment of hypophysectomized rats proved r pGH with high native bioactivity was obtained.

关 键 词：基因工程 猪生长激素 复性技术 复性 重折叠 

分 类 号：Q57[生物学—生物化学]