基因工程猪生长激素的分离提纯及变复性技术的研究  

Isolation, Purification and Renaturation of Recombinant-DNA-Derived Porcine Somatotropin

作　　者：李震[1] 于瑞嵩[1] 张平[1] 王英[1] 刘惠莉[1] 

机构地区：[1]上海市农业科学院畜牧兽医研究所

出　　处：《生物工程学报》2001年第6期703-705,共3页Chinese Journal of 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 ( 9843190 0 2 )~~

摘　　要：Large scale abstraction and isolation of bacterially synthesized, recombinant-DNA-derived, porcine growth hormone (r-pST) is described. The r-pGH is found in genetic engineering E.coli as the form of inclusion bodies. Pellet fraction which were mainly inclusion bodies, after cell breakage and centrifugation, were collected. Cell envelope components, such as protein, lipid, endotoxin and nucleic acids are selectively removed from the pellet fraction by an EDTA/lysozyme/deoxycholate extraction. Inclusion bodies were dissolved using 6mol/L guanidine/HCl and air oxidation is then carried out in the presence of the guanidine/HCl. The Guanidine/HCl protein mixture were diluted by renaturation solution. Guanidine/HCl were removed by dialysis and then correctly refolded, oxidized r-pGH were obtained. Injection experiment of hypophysectomized rats proved r-pST with high native bioactivity was obtained.Large scale abstraction and isolation of bacterially synthesized, recombinant-DNA-derived, porcine growth hormone (r-pST) is described. The r-pGH is found in genetic engineering E.coli as the form of inclusion bodies. Pellet fraction which were mainly inclusion bodies, after cell breakage and centrifugation, were collected. Cell envelope components, such as protein, lipid, endotoxin and nucleic acids are selectively removed from the pellet fraction by an EDTA/lysozyme/deoxycholate extraction. Inclusion bodies were dissolved using 6mol/L guanidine/HCl and air oxidation is then carried out in the presence of the guanidine/HCl. The Guanidine/HCl protein mixture were diluted by renaturation solution. Guanidine/HCl were removed by dialysis and then correctly refolded, oxidized r-pGH were obtained. Injection experiment of hypophysectomized rats proved r-pST with high native bioactivity was obtained.

关 键 词：rPGH 分离提纯 变复性 基因工程猪生长激素 

分 类 号：Q575.11[生物学—生物化学] Q789