厦门大学学报（自然科学版）2016，Vol.55Issue(2)：192-197,6.DOI:10.6043/j.issn.0438-0479.2016.02.008
重组人胸腺肽α1在大肠杆菌中的表达纯化与鉴定
Expression,Purification and Identification of Recombinant Human Thymosin-alpha 1 in Escherichia coli
摘要
Abstract
In the present study,expression,purification and identification of recombinant human thymosin-alpha 1 (rhTα1)in Esche-richia coli were reported.Firstly,bacteria expressing the rhTα1 fusion protein were utilized in the 300 L fermentator,and protein was induced by IPTG.Then,bacteria were harvested by centrifugation,and cell lysate was obtained using high pressure crushes.The crude protein was purified using the affinity chromatograghy,and the yield of the fusion protein was up to 170 mg per 1 L culture. Furthermore,the crude protein was processed by enterokinase (EK)proteolysis,affinity chromatography,reversed phase chromatog-raphy and exclusion chromatography to obtain the high purity of rhTα1 .SDS-PAGE and HPLC-RPC analysis indicated that rhTα1 could reach the purity of more than 99%.Additionally,rosette formation assay demonstrated that the activity of rhTα1 was quite similar to that of the commercial standard product.Molecular weight of rhTα1 by MALDI-TOF MS was 3066.59 u,which agreed with the theoretic result (3066 u),and the analysis of N-terminal of rhTα1 showed an accordance with the theoretic data,which all implied a potential for the industrialization of rhTα1 .关键词
重组人胸腺肽α1(rhTα1)/表达/纯化Key words
recombinant human thymosin-alpha 1 (rhTα1)/expression/purification分类
生物科学引用本文复制引用
尹凤红,肖清江,周卫东,陈清西..重组人胸腺肽α1在大肠杆菌中的表达纯化与鉴定[J].厦门大学学报（自然科学版）,2016,55(2):192-197,6.基金项目
国家高技术研究发展计划(863)项目(2007AA021604) （863）
