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胸腺肽α_1的温度诱导原核可溶性表达与简易纯化 被引量：3

Thermo-inductively Prokaryotic Expression in a Soluble Form and Easy Purification of Thymosin α_1
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摘要 将人工合成的人胸腺肽α1(Tα1)基因插入到载体pBV222,转化大肠杆菌DH5α菌株并置42℃诱导表达可溶性融合蛋白。融合蛋白经镍亲和层析柱纯化,加入肠激酶切割,再将酶切反应体系回到镍亲和层析柱。所获穿过液中的重组胸腺肽α1(rTα1),经十二烷基硫酸钠-聚苯烯酰胺凝胶电泳(SDS-PAGE)和反相高效液相色谱(RP-HPLC)分离及分析,纯度分别为94.5%和72%。以癌症患者外周血淋巴细胞开展玫瑰花环试验,所获rTα1纯品显示出与化学合成Tα1(sTα1)相同的生物学活性。 An artificially synthesized gene of human thymosin α1（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 dodecyl 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.
作者 陈培富 李红芳 鲁琼芬 陈俊
机构地区 云南农业大学动物科学技术学院
出处 《云南农业大学学报（自然科学版）》 CAS CSCD 北大核心 2012年第2期183-187,共5页 Journal of Yunnan Agricultural University:Natural Science
基金 云南农业大学博士科研启动经费资助项目(A2002094)
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 thymosin α1 prokaryotic expression system nickel affinity chromatography reverse phase high performance liquid chromatography rosette test
分类号 Q78 [生物学—分子生物学]
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参考文献22

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二级参考文献42

共引文献31

同被引文献40

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引证文献3

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云南农业大学学报（自然科学版）
2012年 第2期

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