pIRES2-EGFP-prnd真核表达载体的构建及其在BHK细胞中的表达

摘要

为探讨叠朊蛋白(Doppel)在正常生理条件及相关疾病中的临床及生物学意义,构建BALB/c小鼠Doppel基因prnd的重组真核表达质粒pIRES2-EGFP-prnd,并转染BHK细胞,IFA检测其表达情况.结果表明,Doppel蛋白通过pIRES2-EGFP-prnd在BHK细胞中表达.试验可为进一步研究Doppel蛋白的结构、正常的生理生化功能及其与相关疾病的关系提供重要工具.%To investigate the clinical and biological significance of Doppel in normal physiological conditions and related diseases,the prnd gene of the testis of BALB/c mouse was cloned into the eukaryotic expression vector pIRES2-EGFP.The positive recombinant was designated with pIRES2-EGFP-prnd.After being identified by restriction enzyme digestion and sequencing,the recombinant pIRES2-EGFP-prnd was transfected into BHK cells by liposomes.The expression of the recombinant pIRES2-EGFP-prnd in the BHK cells was observed by fluorescence microscope and IFA.The result showed that the prnd gene was successfully expressed in the BHK cells.It will provide an important tool to further study the structure of Doppel protein,its normal physiological and biochemical functions and its relationship with related diseases.