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| 酵母双杂合系统AD端阴离子交换蛋白C-末端表达质粒的构建 | |
| 引用本文： | 李宏涛,傅国辉,秦勇,杜洪清,姜晓姝,刘明,孔宪刚. 酵母双杂合系统AD端阴离子交换蛋白C-末端表达质粒的构建[J]. 生物医学工程学杂志, 2002, 19(2): 274-286,290 |
| 作者姓名： | 李宏涛 傅国辉 秦勇 杜洪清 姜晓姝 刘明 孔宪刚 |
| 作者单位： | 1. 哈尔滨医科大学,病理生理学教研室,哈尔滨,150086 2. 哈尔滨市职工医院,哈尔滨,150041 3. 中国农业科学院,哈尔滨兽医研究所,生物技术室国家重点实验室,哈尔滨,150001 |
| 基金项目： | 国家自然科学基金资助项目 (3 9970 2 91) |
| 摘 要： | 利用PCR方法，从阴离子交换蛋白1（AE1）全长cDNA中扩增出约350bp c末端cDNA片段，测序后将其克隆至pGADT7载体上，用醋酸锂法构建好的pADT7-AE1-c末端转染酵母菌HA109，观察其在选择性培养基上的表达情况。结果表明，获得了530bp AE1c－末端cDNA,pGADT7-AE1-c末端对酵母无毒性，不能激活检测基因，可作为酵母双杂合系统中的靶基因。 |
| 关 键 词： | 酵母双杂合系统 AD端阴离子交换蛋白 C－末端 质粒 构建 克隆 |
Cloning of AE1-c-end cDNA and Construction of Its Expression Plasmid for Yeast Two-Hybrid System |
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| Li Hongtao Fu Guohui Qin Yong Du Hongqing Jiang Xiaoshu Liu Ming Kong Xiangang. Cloning of AE1-c-end cDNA and Construction of Its Expression Plasmid for Yeast Two-Hybrid System[J]. Journal of biomedical engineering, 2002, 19(2): 274-286,290 | |
| Authors: | Li Hongtao Fu Guohui Qin Yong Du Hongqing Jiang Xiaoshu Liu Ming Kong Xiangang |
| Affiliation: | Department of Pathophysiology, Harbin Medical University, Harbin 150086. |
| Abstract: | In this study, about 350 bp cDNA fragment was amplified by PCR. After being sequenced, the AE1-c-end gene fragment was cloned into EcoR I-Pst I site of pGADT7 to form AD ends in the yeast two-hybrid system. The recombinant plasmid was transformed into yeast AH109, and the expression in the yeast was observed. The results demonstrate that AE1-c-end was obtained. pGADT7-AE1-c-end has no toxic effect on the yeast. It can serve as a target gene of yeast two-hybrid system. |
| Keywords: | Anion Exchanger 1 Yeast two hybrid system Clone |
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