云南农业大学学报2012，Vol.27Issue(2)：183-187,5.DOI:10.3969/j.issn.1004-390X(n).2012.02.008
胸腺肽α1的温度诱导原核可溶性表达与简易纯化
Thermo-inductively Prokaryotic Expression in a Soluble Form and Easy Purification of Thymosin α1
摘要
Abstract
An artificially synthesized gene of human thymosin a, (Tα1) was inserted into pBV222 and then thermo-inductively expressed as a fusion protein in a soluble form in transformed Escherichia coli strain DH5α at 42°C. The fusion protein purified through nickel affinity chromatography was cleaved with enterokinase. The cleavage mixture was returned to the nickel affinity chromatography column. Recombinant Tα1 (rTα1) in the flow-through reached a purity of 94. 5% as showed by sodium dode-cyl sulfate-polyacrylamide gel electrophoresis ( SDS-PAGE ) and 72% by reverse phase-high performance liquid chromatography (RP-HPLC), respectively. The finally purified rTα1, exhibited the same bioactivity as chemically synthesized Tα1, in rosette tests with peripheral blood lymphocytes from cancer patients.关键词
胸腺肽α1/原核表达系统/镍亲和层析/反相高效液相色谱/玫瑰花环试验Key words
thymosin α1/prokaryotic expression system/nickel affinity chromatography/reverse phase high performance liquid chromatography/ rosette test分类
生物学引用本文复制引用
陈培富,李红芳,鲁琼芬,陈俊..胸腺肽α1的温度诱导原核可溶性表达与简易纯化[J].云南农业大学学报,2012,27(2):183-187,5.基金项目
云南农业大学博士科研启动经费资助项目(A2002094). （A2002094）
