Using The Yeast Two-hybrid System With The Fmrp-interacting Proteins

Fragile X syndrome is the most common cause of inherited mental retardation whose main clinical features are moderate mental retardation, enlarged testes and typical facies with large ears, high forhead and prominent lower jaw. Down-regulation of its disease gene — fragile mental retardation gene 1 ( FMR1 ) results in various clinical manifestations.The function of its protein product FMRP has been the focus of fragile X syndrome study and much progress has been made by different studies, for example, by study of FMR1 gene expression regulation, interaction of FMRP with RNA, alternative splicing of FMR1 gene expression, interaction of FMRP with other proteins, localization of FMRP in cells. But uptill now, the biological role of FMRP , or the cell process FMRP involves in, is still not known though FMRP is postulated to involve in mRNA metabolism or translation regulation. In an attempt to find new clues to the function of FMRP, we try to find the proteins interact with FMRP by using the yeast two-hybrid system.The yeast two-hybrid system is a powerful tool to study protein-protein interaction in which two interacting proteins are fused separately to the DNA-binding domain and transcription activation domain of transcription factors and the interaction between the two proteins are monitored in yeast by the expression status of report genes. By this method, DNAs of proteins interact with target protein can be directly obtained.This work is divided into the following two parts:1. Interaction of six natural FMRP isoforms with FXR1 proteinFXR1 and FXR2 are two genes highly homologous to FMR1, whose proteins interact with FMRP. The interaction region is localized inside exon 7...