Of Fxr1p The Interaction Of Rna Library Screening

Objective: Fragile X syndrome (FXS) is one of the most common form of inherited cognitive impairment, which have a diversity of clinical manifestation. The mutation of FMR1 causes FXS. FXR1, a gene that shares great similarity with FMR1, (Fragile X related gene 1) is a housekeeping gene that plays an important role in the development process of nervous system and muscular system. FXR1P(Fragile X related protein 1), the translational product of FXR1, is a kind of RNA-binding protein, but our knowledge about FXR1P-targeting RNAs is extremely limited. Therefore, we screened from a human brain gyrus hippocampi pRH3-cDNA library for FXR1P-interacting RNAs in order to enrich our knowledge about the function of FXR1.Methods: Construct human braingyrus hippocampi pRH3-cDNA library. The FXR1P whole long coding vector pYESTrp3/ FXR1 was introduced into host strain L40 ura3/ pHybLex/ Zeo-MS2;Then the toxicity and self-activation of pYESTrp3/ FXR1 were tested; The human brain library was screening by yeast transformation; The plasmids were extracted from the obtained positive yeast clones and reintroduced into yeast strain L40 ura3/ pHybLex/ Zeo-MS2/ pYESTrp3/ FXR1 one by one so as to validate the RNA-protein interactions; The insert fragments of identified positive clones were sequenced and analyzed with BLAST in NCBI.Results: The human braingyrus hippocampi pRH3-cDNA library was successfully constructed; The introduction of pYESTrp3/ FXR1 was harmless to the host strain and there was no self-activation; 56 strains of positive results were obtained through yeast three hybrid screening and nine of them were verified by one-to-one yeast three hybrid system analysis; The nine clones were sequenced and then conformed by BLAST analysis; four of them proved to be inserted correctly, namely, Homo sapiens tubulin, alpha, ubiquitous (K-ALPHA-1),Homo sapiens IQ motif containing E (IQCE),Homo sapiens dipeptidyl-peptidase 7(DPP7),Homo sapiens leukocyte receptor cluster(LRC)member 4, while five of them were inserted reversely. Our findings may cast a new light on FXR1P function.Conclusions: A platform of the yeast three-hybrid system was established to perform research on RNA-protein interaction; Constructed the human braingyrus hippocampi pRH3-cDNA library; Four correctly inserted gene fragments were identified whose mRNAs may interact with FXR1P.