文献类型：期刊文献

中文题名：TRAIL包涵体在细胞内重折叠现象的初步研究

英文题名：Preliminary studies on in vivo refolding of TRAIL inclusion bodies

作者：汤亚南[1];潘若鋆[1];周劲松[2];张国钧[2];孙菁[2];球谊[2];魏东芝[1];沈亚领[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室鲁华生物技术研究所,上海200237;[2]上海恰尔生物技术有限公司,上海201103

年份：2009

卷号：39

期号：4

起止页码：32

中文期刊名：工业微生物

外文期刊名：Industrial Microbiology

收录：CSCD:【CSCD2011_2012】；

基金：国家863重大科技专项滚动项目(2004AA223801);上海市重点学科建设项目(B505)资助

语种：中文

中文关键词：肿瘤坏死因子相关凋亡诱导配体;重折叠;包涵体;重组大肠杆菌

外文关键词：Apo2L/TRAIL; inclusion bodies; in vivo refolding; recombinant Escherichia coli

摘要：肿瘤坏死因子相关凋亡诱导配体(Apo2L/TRAIL)是一种新型的抗肿瘤蛋白。但在大肠杆菌表达系统中,表达的TRAIL蛋白往往容易聚集形成包涵体。本文对重组TRAIL包涵体在胞内重折叠转化成可溶性TRAIL蛋白作了初步探索,结果表明:当蛋白表达后,迅速降低温度至30℃并添加50μg/mL氯霉素继续培养4 h,可溶性TRAIL产量可提高至16.7%。这一结果在3.7L罐上也得到了证实,可溶性TRAIL表达量达到14.5%。
TRAIL as a novel anticancer therapeutic agent in development, however, is prone to aggregate into inclusion bodies when expressed in recombinant Escherichia coll. The applicability of in vivo refolding of inclusion bodies was well evaluated. The results showed that after induction, the yield of soluble TRAIL was significantly enhanced to 16.7 % when cultivated further at 30 ℃ for 4 h, prior to arrest of protein synthesis by chloramphenicol addition （up to 50μg/mL）. It was confirmed successfully in 3.7 L fermentor for producing 14.5 % soluble TRAIL.