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Cloning, expression, purification,identification and biological activity of recombinant human thymosin beta 4
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 16
- DOI:
- 10.3321/j.issn:1000-2790.2008.16.004
- Key Word:
- 胸腺素β4;串联重复序列;克隆,分子;基因表达
Abstract： 目的:利用基因工程的方法原核表达重组人胸腺素β4(Tβ4)并对其进行纯化和鉴定. 方法:使用大肠杆菌偏爱密码子合成人Tβ4全长基因,构建了Tβ4的二串联体基因(Tβ4②),将其克隆入表达载体pET-22b(+)中,重组质粒转化大肠杆菌BL21(DE3)感受态,IPTG诱导表达后,经盐析、疏水层析和离子交换层析纯化重组蛋白,采用免疫印迹和基质辅助激光解吸电离飞行时间质谱对重组蛋白进行鉴定. 结果:获得了纯化的重组Tβ4②蛋白,并具有生物学活性. 结论:成功构建、表达和纯化了Tβ4②,为其功能研究奠定基础.
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