Position: Home > Articles > Study on renaturation of recombinant bovine interleukin-2 expressed in Escherichia coli
Acta Agriculturae Shanghai
2006,22
(4)
27-32
大肠杆菌表达重组牛IL-2复性技术的研究(英文)
作 者：
黄建珍;于瑞嵩;沈秋姑;李祥瑞;李震
单 位：
上海市农业科学院畜牧兽医研究所;南京农业大学动物医学院;江西农业大学动物科技学院
关键词：
重组牛白细胞介素2;包含体;复性
摘 要：
研究了重组牛白介素2复性条件,如pH,蛋白浓度,DTT浓度,氧化剂(GSSG)和还原剂(GSH)的比例,Cu2+及L-Arg,低浓度的盐酸胍的添加,复性方法等。结果表明,复性液中添加20 mmol/mL DTT可以提高rBoIL-2的复性效率;利用尿浓度梯度复性法3.5 mg/mL的rBoIL-2蛋白溶液复性率可达50%以上,测定活性效价达9.6×104IU/mL。
译 名：
Study on renaturation of recombinant bovine interleukin-2 expressed in Escherichia coli
作 者：
Huang Jian-zhen~1,3,YU Rui-song~1,2*,SHEN Qiu-gu~3,LI Xiang-rui~4,LI Zhen~1,2**(~1Animal Husbandry and Veterinary Research Institute,Shanghai Academy of Agricultural Sciences,Shanghai 201106,China;~2 Shanghai Municipal Key Laboratory of Agri-Genetics and Breeding,Shanghai 201106,China;~3 College of Animal Science,Jiangxi Agricultural University,Nanchang 330045,China;~4 Faculty of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)
关键词：
Recombinant bovine interleukin-2(rBoIL-2);Inclu
摘 要：
Recombinant bovine interleukin-2 forms insoluble inclusion body when overexpressed in Escherichia coli, and must be denatured and renatured in vitro before acquiring biological activity.We systematically studied such conditions of renaturing rBoIL-2 as pH,protein concentration,DTT concentration,ratio of oxidizer(GSSG) to reducer(GSH),and additions of Cu~2+,L-Arginine and Guandine-HCl of low concentration,as well as renaturation methods.The results showed that adding 20 mmol/L DTT to denaturation solution could improve the efficiency of the subsequent rBoIL-2 renaturation,and the refolding rate of 3.5 mg/mL rBoIL-2 protein solution could reach 50% by means of urea gradient renaturation,in which the bioactivity of 9.6×10~4 IU/mL could be obtained.
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