| 人CDK2-AP1(DOC-1)酵母双杂交诱饵质粒自激活作用鉴定 |
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| 引用本文： | 倪前伟,孙沫逸,戴建武,韩津,曹立雪,金伟,郑军,李建虎.人CDK2-AP1(DOC-1)酵母双杂交诱饵质粒自激活作用鉴定[J].北京口腔医学,2010,18(1):5-7,17. |
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| 作者姓名： | 倪前伟 孙沫逸 戴建武 韩津 曹立雪 金伟 郑军 李建虎 |
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| 作者单位： | 第四军医大学口腔医学院口腔颌面-头颈肿瘤外科,西安,710032;中国科学院遗传与发育生物学研究所 |
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| 基金项目： | 国家自然科学基金资助项目(30572059);;陕西省科技攻关项目[2006K09-G10(4)];;教育部留学人员科研启动基金(2002HG001) |
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| 摘 要： | 目的验证诱饵质粒pBD—DOC-1在酵母双杂交系统的自激活性及毒性作用，为应用酵母双杂交系统（yeasttwo—hybrid system）筛选与p12DOC-1／CDK2-AP1相互作用的蛋白建立实验基础。方法将诱饵质粒pBD—DOC-1转化到酵母细胞MAV203中，检测诱饵蛋白有无毒性和自激活作用。同时利用对照质粒组筛选组氨酸（His）本底表达抑制剂3AT（3-氨基．1，2，4-三唑）的合适工作浓度。结果诱饵质粒pBD．DOC—1成功转化到酵母细胞MAV203中，对宿主酵母细胞无毒性，对报告基因无自激活作用。确定了组氨酸（His）本底表达抑制剂3AT（3-氨基-1，2，4-三唑）的合适工作浓度。结论诱饵质粒pBD—DOC-1可以用于酵母双杂交实验，为进-步运用酵母双杂交技术在人类组织cDNA文库中筛选与之相互作用的蛋白奠定了基础。
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| 关 键 词： | p12DOC-1/CDK2-AP1 酵母双杂交 自激活 |
Identification of a yeast two-hybrid bait vector of p12~(DOC-1/CDK2-AP1) |
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| NI Qian-wei,SUN Mo-yi,DAI Jian-wu,HAN Jin,CAO Lixue,JIN Wei,ZHENG Jun,LI Jian-hu.Identification of a yeast two-hybrid bait vector of p12~(DOC-1/CDK2-AP1)[J].Beijing Journal Of Stomatology,2010,18(1):5-7,17. |
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| Authors: | NI Qian-wei SUN Mo-yi DAI Jian-wu HAN Jin CAO Lixue JIN Wei ZHENG Jun LI Jian-hu |
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| Institution: | NI Qian-wei,SUN Mo-yi,DAI Jian-wu,HAN Jin,CAO Li-xue,JIN Wei,ZHENG Jun,LI Jian-hu.Department of Oral , Maxillofacial Surgery,Stomatological College,Fourth Military Medical University,Xi'an 710032,China |
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| Abstract: | Objective To identify the self-activation and the toxic effects of bait plasmid pBD-DOC-1 in yeast two-hybrid system and to screen the protein that interacts with p12DOC-1/CDK2-AP1.Methods The bait plasmid pBD-DOC-1 was transformed into yeast cells MAV203,and the toxicity and self-activation of bait protein detected.Meantime,taking advantage of the control plasmid,we selected the appropriate working concentration of 3AT (3-amino-1,2,4-triazole) which could inhibit the background expression of his amino acid... |
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| Keywords: | p12DOC-1/CDK2-AP1 Yeast two hybrid system Self-activation |
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