影响基因工程猪生长激素复性效率的因素  

THE FACTORS AFFECTING RENATURATION EFFICIENCY OF RECOMBINANT PORCINE GROWTH HORMONE

作　　者：于瑞嵩[1,2] 李震[1,2] 刘惠莉 张平[1,2] 张德福 

机构地区：[1]上海市农业科学院畜牧兽医研究所 [2]上海市农业遗传育种重点实验室动物遗传工程分室,上海201106

出　　处：《上海农业学报》2004年第1期26-29,共4页Acta Agriculturae Shanghai

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 (9843 190 0 2 )资助

摘　　要：对影响基因工程猪生长激素 (r pGH)体外复性效率的因素进行了研究 ,结果表明 :复性液组成、变性剂种类、复性方法的选择、变复性pH对r pGH体外复性效率有较大影响 ,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。提取包含体用 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 ,采用透析复性法———复性缓冲液Ⅲ (0 .2 5 %NaHCO3,0 .2 %α 乳糖 ,0 .2 %甘露醇 ) ,pH9.0 ,透析 5～ 6次 ,可较好地恢复r pGH生物活性。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 factors affecting in vitro renaturation efficiency of r-pGH were studied. The results showed that the variety of denaturant, the composition and pH of renaturation solution, the kind of methods selected for renaturation had significant effects on the in vitro renaturation efficiency of r-pGH; but the addition of 2-mercaptoethanol and GSH or GSSG had less effects. The inclusion bodies extracted from host cells were dissolved using 6mol/L guanidine/HCl and air oxidation for 80h was carried out in the presence of guanidine/HCl. The guanidine/HCl protein mixture was then diluted by renaturation solution. Guanidine/HCl was removed by dialysis against renaturation solution Ⅲ(0.25% NaHCO 3, 0.2% lactose, 0.2% mannitol, pH9.0) for 5 to 6 times and correctly refolded, oxidized r-pGH was 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 复性效率 影响因素 

分 类 号：S828[农业科学—畜牧学]