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Expression of Bm-TFF2 mutants in Escherichia coli andtheir cell migration-promoting activity

yu guo-Yu XIANG Yang ZHANG Hong-Yun JIANG Ping Lee Wen-Hui ZHANG Yun ZHANG

yu guo-Yu, XIANG Yang, ZHANG Hong-Yun, JIANG Ping, Lee Wen-Hui, ZHANG Yun, ZHANG. Expression of Bm-TFF2 mutants in Escherichia coli andtheir cell migration-promoting activity. Zoological Research, 2011, 32(4): 379-385. doi: 10.3724/SP.J.1141.2011.04379
Citation: yu guo-Yu, XIANG Yang, ZHANG Hong-Yun, JIANG Ping, Lee Wen-Hui, ZHANG Yun, ZHANG. Expression of Bm-TFF2 mutants in Escherichia coli andtheir cell migration-promoting activity. Zoological Research, 2011, 32(4): 379-385. doi: 10.3724/SP.J.1141.2011.04379

三叶因子Bm-TFF2突变体在原核体系中的表达及促细胞迁移活性

doi: 10.3724/SP.J.1141.2011.04379
基金项目: 中国科学院“西部之光”(Y102291081); “973”项目(2010CB529800); 国家基金委面上项目(30870304)
详细信息
    作者简介:

    余果宇(1972), 女, 博士,研究方向为胃肠肿瘤的分子生物学; 向阳(1983-), 男, 博士生,研究方向为胃肠肿瘤的细胞生物学

    向 阳:余果宇(1972), 女, 博士,研究方向为胃肠肿瘤的分子生物学; 向阳(1983-), 男, 博士生,研究方向为胃肠肿瘤的细胞生物学

    通讯作者:

    张 勇

Expression of Bm-TFF2 mutants in Escherichia coli andtheir cell migration-promoting activity

More Information
    Author Bio:

    yu guo-Yu,XIANG Yang

    yu guo-Yu,XIANG Yang

    Corresponding author: ZHANG Yong
  • 摘要: 大蹼铃蟾三叶因子Bm-TFF2具有较人TFF2更强的促细胞迁移和抗凋亡活性。该研究利用RT-PCR方法扩增得到野生型Bm-TFF2的基因, 然后分别构建N端、C端和分子中两个精氨酸突变的突变体, 最后连接表达载体产生pET32a(+)/Bm-TFF2突变型重组质粒, 转入大肠杆菌中, 经37 °C培养, IPTG诱导, 其融合蛋白主要存在于包涵体中, 用组氨酸标签的亲合柱纯化溶解后的包涵体上清, 进一步用RP-HPLC纯化得到硫氧还蛋白(TRX)/Bm-TFF2突变型融合蛋白。通过SDS-PAGE和Western blotting检测分析其纯度和特异性。最终, 从1L培养基中得到20 mg纯度为95%的三种重组突变型融合蛋白。三种突变型重组蛋白都具有剂量依赖性的促细胞迁移活性, 并且其活性无显著差异。该研究为进一步研究Bm-TFF2结构和功能的关系以及揭示其作用的分子机制奠定了基础。
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出版历程
  • 收稿日期:  2011-01-05
  • 修回日期:  2011-05-18
  • 刊出日期:  2011-08-22

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