DLX4基因酵母双杂交系统的构建及自激活检测

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

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M aV203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。

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.