FKBP25缺失突变体在细胞中的表达与定位  

Localization and expression of human FKBP25 deletion mutants in mammalian cell lines

作　　者：张姗靖[1] 王蓓华[2] 龚芮[2] 潘林鑫[2] 耿慧武[2] 刘晓颖[2] 邓松华[1] 范礼斌[2] 

机构地区：[1]安徽医科大学生命科学学院生物学教研室,合肥230032 [2]安徽医科大学病理生理学教研室生命科学学院生物学教研室,合肥230032

出　　处：《安徽医科大学学报》2014年第9期1250-1253,共4页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:11040606M170、11040606M164)

摘　　要：目的研究FKBP25缺失突变体蛋白在细胞内的表达和定位。方法以人FKBP25全长cDNA序列的质粒为模板,PCR方法分别扩增出FKBP25缺失突变体序列,然后插入真核表达载体pcDNA3.1中,通过Western blot方法和免疫荧光方法检测其在细胞株中的表达和定位。结果成功构建了FKBP25缺失突变体的真核表达载体,经Western blot检测FKBP25缺失突变体在HEK 293T细胞中能够有效表达,免疫荧光显微镜结果显示FKBP25缺失突变体在COS7细胞中分布于细胞核和细胞质。结论 FKBP25缺失突变体在HEK 293T、COS7细胞中能够表达,为了解FKBP25缺失突变体的功能提供了一定基础。Objective To investigate the expression and localization of the product of FKBP25 deleted mutants in cells. Methods The full length eDNA fragment of FKBP25 was used for PCR amplification, to construct FKBP25 deletion mutant gene sequence respectively, and inserted them into pcDNA3.1 vector. All expressions were detected by Western blot and immunofluorescenee. Results The expression of FKBP25 deletion mutants was constructed successfully. Western blot assay showed that FKBP25 deletion mutants could express efficiently in HEK293T cell line, and the results of immunofluorescence microscopy showed that the FKBP25 deletion mutants protein was found to be localized in the nucleus and cytoplasm in COS7 cells lines. Conclusion The FKBP＇25 deletion mutants can express in HEK 293T and COS7 cell lines. The results provide the basis for further understanding of the function of FKBP25 deletion mutants.

关 键 词：FKBP25 基因表达 免疫荧光 细胞定位 

分 类 号：R341[医药卫生—基础医学] R394.2