利用酵母双杂交系统筛选与FXR1P相互作用的蛋白

Screening of proteins binding to FXR1P using yeast two-hybrid technique

目的筛选FXR1P相互作用蛋白,探讨FXR1P生物学功能。方法应用酵母双杂交技术,构建诱饵表达载体pGBKT7/FXR1,转化酵母菌AH109,与转化了人胎脑cDNA文库的酵母菌Y187交配筛选阳性克隆并测序。结果重组表达载体pGBKT7/FXR1构建成功;从人胎脑cDNA文库中筛选出5个与FXR1P结合的蛋白基因,分别与5种已知的编码蛋白基因序列同源:胞苷一磷酸-N-乙酰神经氨酸合成酶(CMAS)、铁蛋白重链多肽1(FTH1)、高尔基体自身抗原golgina亚类4(GOLGA4)、17-β羟类固醇脱氢酶1(HSD17B1)、绒毛膜生长催乳激素1(CSH1)。结论为进一步研究FXR1P的功能及脆性X综合征发病机制提供新的线索。

Objective To screen the proteins interacting with FXR1P for functional investigation of FXR1P. Methods The yeast strain AH109 transformed with the recombinant expression vector pGBKT7/FXR1 was mated with the yeast strain Y187 pretransformed with human fetal brain cDNA library. The positive clones were screened and identified by sequence analysis. Results The recombinant expression vector pGBKT7/FXR1 was constructed successfully. Five proteins binding to FXR1P were screened from human fetal brain cDNA library using the yeast two-hybrid system, including CMAS, FTH1, GOLGA4, HSD17B1 and CSH1. Conclusion These results provide new clues for investigating the biological functions of FXR1P and the pathogenesis of Fragile X syndrome.