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Expression and identification of Bm-TFF2 in Pichia pastoris

Yu guo-yu ZHANG Hong-Yun ZHANG Yong JIANG Ping Lee Wen-Hui Zhang Yun

Yu guo-yu, ZHANG Hong-Yun, ZHANG Yong, JIANG Ping, Lee Wen-Hui, Zhang Yun. Expression and identification of Bm-TFF2 in Pichia pastoris. Zoological Research, 2010, 31(6): 565-569. doi: 10.3724/SP.J.1141.2010.06565
Citation: Yu guo-yu, ZHANG Hong-Yun, ZHANG Yong, JIANG Ping, Lee Wen-Hui, Zhang Yun. Expression and identification of Bm-TFF2 in Pichia pastoris. Zoological Research, 2010, 31(6): 565-569. doi: 10.3724/SP.J.1141.2010.06565

Bm-TFF2在毕赤酵母中的表达及鉴定

doi: 10.3724/SP.J.1141.2010.06565
基金项目: “973” 项目(2010CB529805); 国家基金委重点项目(30630014); 国家基金委面上项目(30570359); 中国科学院重要方向项目(KSCX2- YW-R-088)
详细信息
  • 中图分类号: Q966; Q516; Q786

Expression and identification of Bm-TFF2 in Pichia pastoris

  • 摘要: Bm-TFF2, 一种从大蹼铃蟾皮肤分泌物中分离得到的两栖类三叶因子, 具有比人三叶因子更强的生物学活性。该研究以Bm-TFF2的cDNA为模板, 利用PCR方法扩增Bm-TFF2基因, 然后插到含有AOX1启动子和α-因子信号肽序列的表达载体pPIC9K中, 采用毕赤酵母表达系统进行分泌表达, 并用G418筛选高拷贝整合转化子。SDS-PAGE和Western blotting都检测到Bm-TFF2被分泌性表达于酵母上清。在1%的甲醇诱导表达不同时间后, 重组蛋白在72 h的表达量最大, 可达50 mg/ L; 而不同浓度的饱和硫酸铵沉淀菌液上清时, 80%的饱和硫酸铵沉淀量最大。这些结果表明, 重组质粒Bm-TFF2-pPIC9K成功构建并在真核细胞中高效表达, 这为进一步研究Bm-TFF2的生物学活性及其结构与功能关系奠定了基础。
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出版历程
  • 收稿日期:  2010-08-17
  • 修回日期:  2010-11-10
  • 刊出日期:  2010-12-22

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