人肠三叶因子酵母双杂交载体的构建及其自激活作用鉴定
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卢雅丕, 董 菁, 周 飞, 王 琳, 陈美娅, 廉亚美, 张 波, 任建林
肠三叶因子;酵母双杂交载体;自激活作用,卢雅丕,董菁,周飞,王琳,陈美娅,廉亚美,张波,任建林,通讯作者:,Constructionofahumanintestinaltrefoilfactoryeasttwo-hybridvectorandid
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卢雅丕, 董菁, 周飞, 王琳, 陈美娅, 廉亚美, 张波, 任建林, 厦门大学附属中山医院消化内科 厦门大学消化疾病研究所 厦门市消化疾病中心 福建省厦门市 361004
福建省自然科学基金资助项目, No. 2006J0380
福建省卫生厅青年科研基金资助, No. 2004-2-6和No. 2006-1-53
通讯作者: 任建林, 361004, 福建省厦门市, 厦门大学附属中山医院消化内科. jianlin.ren@xmzsh.com
电话: 0592-2993170 传真: 0592-2993170
收稿日期: 2007-09-09 修回日期: 2007-11-02
Construction of a human intestinal trefoil factor yeast two-hybrid vector and identification of its self-activation in yeast cells
Ya-Pi Lu, Jing Dong, Fei Zhou, Lin Wang, Mei-Ya Chen, Ya-Mei Lian, Bo Zhang, Jian-Lin Ren
Ya-Pi Lu, Jing Dong, Fei Zhou, Lin Wang, Mei-Ya Chen, Ya-Mei Lian, Bo Zhang, Jian-Lin Ren, Department of Gastroenterology, Zhongshan Hospital, Xiamen University; Gastroenterology Institute of Xiamen University, Gastroenterology Center of Xiamen, Xiamen 361004, Fujian Province, China
Supported by: the Natural Science Foundation of Fujian Province, No. 2006J0380 and Science Fund for Young Scholars of Department of Health Fujian Province, No. 2004-2-6, No. 2006-1-53
Correspondence to: Jian-Lin Ren, Department of Gastroenterology, Zhongshan Hospital, Xiamen University, Xiamen 361004, Fujian Province, China. jianlin.ren@xmzsh.com
Received: 2007-09-09 Revised: 2007-11-02
Abstract
AIM: To construct a yeast two-hybrid bait vector of intestinal trefoil factor (ITF/TFF3) and identify its self-activation.
METHODS: The open reading frame of ITF gene containing NcoⅠandXhoⅠ
endoenzyme sites was obtained by RT-PCR from normal human intestinal mucosa and ligated into pGEMT vector from a TA clone. The hTFF3 gene was then transferred into the vector pENTR11 by enzyme digestion and ligation to obtain the entry clone pENTR11-hTFF3. Subsequently, pDEST32-hTFF3, the yeast two-hybrid bait vector of hTFF3, was constructed by the LR reaction. Finally, pDEST32-hTFF3 and pDEST22 were transfected into MaV203 yeast and amplified by yeast in 3AT plates with SC/-leu/-trp/-his ......
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