IQCE mRNA与FXR1P在酵母三杂交体系中的相互作用

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

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。

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 eDNA insert at last. The interest eDNA 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.