IQCE mRNA与FXR1P在酵母三杂交体系中的相互作用

脆性X相关基因1编码蛋白FXR1P是一种RNA结合蛋白,其所结合的靶RNA目前所知甚少。本研究应用酵母三杂交技术对本课题组从pRH3-cDNA人脑海马RNA表达文库中筛选到的一种可能与FXR1P存在相互作用的RNA IQCE进行研究,以验证该RNA与FXR1P的相互作用。方法为:提取利用酵母三杂交技术初步筛选得到的酵母阳性克隆的质粒,转化大肠杆菌Top10,利用其质粒不相容性分离插入了目的片段的pRH3′-cDNA质粒,将该质粒转化入含目的基因FXR1的酵母菌株L40-ura3/pHyb lex/Zeo-MS2/pYESTrp3/FXR1,进行一对一的酵母三杂交验证,最后将该片段进行测序。测序结果为IQCE的一段编码序列,而目前尚无研究报导FXR1与IQCE的相互关系。结论:提示FXR1P与IQCE mRNA存在相互作用,IQCE可能是FXR1P发育调控网络组成成员之一。

The Interaction between IQCE mRNA and FXR1P in Yeast Three-Hybrid System

FXR1P is a RNA-binding protein,but our knowledge about FXR1P's targeting-RNAs is extremely limited.We have previously screened for FXR1P-interacting RNAs from a cDNA library,and the objective of this study is to isolate one of the potential FXR1P-interacting RNAs and validate the interaction by one to one yeast three-hybrid system.The yeast plasmids from a positive clone of yeast three-hybrid system were extracted and introduced into E.coli Top10,the plamid pRH3′-cDNA was isolated through E.coli plasmid incompatibility and re-introduced into L40-ura3/pHyblex/ Zeo-MS2/ pYESTrp3/ FXR1 to perform a one to one three-hybrid test;Sequencing the interest cDNA insert at last.The interest cDNA insert was a part of IQCE and there was no report on the interaction between FXR1P and IQCE mRNA.IQCE mRNA may be a new candidate of FXR1P-interacting RNAs.