上海交通大学学报（医学版）

›› 2010, Vol. 30 ›› Issue (12): 1509-.doi: 10.3969/j.issn.1674-8115.2010.12.015

• 论著（基础研究） • 上一篇    下一篇

DLX4基因酵母双杂交系统的构建及自激活检测

乔勤勤, 孙云燕, 陈玲玲, 陆 萌, 丰有吉   

  1. 上海交通大学附属第一人民医院妇产科, 上海 200080
  • 出版日期:2010-12-25 发布日期:2010-12-31
  • 通讯作者: 孙云燕, 电子信箱: sunyy2003@126.com。
  • 作者简介:乔勤勤（1967—）, 女, 副主任医师；电子信箱: yilun200155@sohu.com。
  • 基金资助:

    国家自然科学基金（30600674）

Construction and self-activation of DLX4 in yeast two-hybrid system

QIAO Qin-qin, SUN Yun-yan, CHEN Ling-ling, LU Meng, FENG You-ji   

  1. Department of Obstetrics and Gynecology, The First Pepole's Hospital, Shanghai Jiaotong University, Shanghai 200080, China
  • Online:2010-12-25 Published:2010-12-31
  • Supported by:

    National Natural Science Foundation of China, 30600674

PDF (PC)

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摘要：

目的 构建DLX4酵母双杂交系统并鉴定其自激活作用。方法 构建人胎盘酵母双杂交cDNA文库，并行PCR鉴定；构建3条DLX4诱饵质粒，包括pDEST32-DLX4(4-1C)、pDEST32-DLX4(4-1N)和pDEST32-DLX4(4-2)；将DLX4诱饵质粒转入酵母MaV203菌株，并检测其在酵母细胞中有无自激活作用。结果 PCR证实成功构建了人胎盘酵母双杂交文库；3条DLX4诱饵质粒中，pDEST32-DLX4(4-2)和pDEST32-DLX4(4-1C)没有自激活作用，可用于文库筛选。结论 成功构建了酵母双杂交文库和DLX4诱饵质粒，可用于筛选与DLX4相互作用的蛋白。

关键词: DLX4, 酵母双杂交, 克隆

Abstract:

Objective To construct a yeast-two hybrid system of DLX4 and identify its self-activation. Methods Human placenta yeast-two cDNA bank was constructed, and was identified by PCR. DLX4 bait plasmids including pDEST32-DLX4(4-1C), pDEST32-DLX4(4-1N) and pDEST32-DLX4(4-2) were constructed and transfected into yeast MaV203 cells, and the self-activation effect was detected. Results It was verified by PCR that human placental cDNA bank was successfully constructed. pDEST32-DLX4(4-2) and pDEST32-DLX4(4-1C) had no self-activation effect, and could be used in the yeast two-hybrid system. Conclusion Yeast-two hybrid bank and DLX4 bait plasmids have been successfully constructed, and can be used to screen the protein interacting with DLX4.

Key words: DLX4, yeast two-hybrid, clone