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重组人TGF-β1融合蛋白的克隆、表达、纯化及鉴定 被引量：4

Cloning, expression, purification and identification of recombinant human TGF-β1 fusion protein
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摘要 目的构建人转化生长因子-β1(TGF-β1)基因的原核表达载体,表达并纯化GST-hTGFβ1.方法应用RT-PCR获得人TGF-β1的基因片段,成功构建人TGF-β1原核表达载体,诱导表达人TGF-β1与GST的融合蛋白;应用亲和层析法纯化重组融合蛋白,对纯化产物进行SDS-PAGE电泳分析和Western Blot检测分析.结果成功构建了人TGF-β1重组融合表达质粒pGEX-4T-1-TGF-β1,表达的蛋白经SDS-PAGE电泳分析,在Mr约为39×103处出现了一条新生的蛋白条带,该表达蛋白具有与TGF-β1抗体特异性的结合能力.结论成功构建了人TGF-β1基因的原核表达载体并纯化了该基因的原核表达产物,为下一步的人TGF-β1自体疫苗的研制奠定了基础. AIM： To clone, express and purify human transforming growth factor β1 （TGF-β1） in E. coli. METHODS： A 336 bp of human rhTGF-β1 gene fragment was obtained by RT-PCR and cloned into pGEX-4T-1 vector, a GST fusion expression vector. The plasmid was transformed into E. coli DH5α and induced to express fusion protein GST- rhTGF-β1 with IPTG. The expression of rhTGF-β1 was detected by SDS-PAGE and Western Blot. RESULTS： A novel protein with expected molecular mass was expressed. The expressed product showed a good binding ability to anti-rhTGF-β1 monoclonal antibody. CONCLUSION ： Our successful cloning and expression of rhTGF-β1 gene and purification of rhTGF-β1 protein lay a basis for the production of rhTGF-β1 autovaccine.
作者 姜昌丽 张英起 颜真
机构地区 第四军医大学药学系生物技术中心
出处 《第四军医大学学报》 北大核心 2006年第3期193-195,共3页 Journal of the Fourth Military Medical University
基金 教育部"长江学者和创新团队发展计划"(IRT0459)
关键词 转化生长因子-Β1 重组融合蛋白/分离与纯化 pGEX4T-1载体 transforming growth factor-β1 recombinant fusionproteins / isolation ＆ purification pGEX-dT-1vector
分类号 Q78 [生物学—分子生物学]
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参考文献8

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共引文献1

同被引文献31

  • 1吴孔田,张英起,颜真.趋化因子受体CCR5胞外段重组蛋白的克隆、表达、纯化及鉴定[J].第四军医大学学报,2006,27(5):412-415. 被引量：3
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第四军医大学学报
2006年 第3期

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