CAJ | 학술논문

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。
목적연구FKBP25결실돌변체단백재세포내적표체화정위。방법이인 FKBP25전장 cDNA서렬적질립위모판， PCR방법분별확증출FKBP25결실돌변체서렬，연후삽입진핵표체재체pcDNA3.1중，통과Western blot방법화면역형광방법검측기재세포주중적표체화정위。결과성공구건료FKBP25결실돌변체적진핵표체재체，경Western blot검측FKBP25결실돌변체재HEK 293T세포중능구유효표체，면역형광현미경결과현시 FKBP25결실돌변체재COS7세포중분포우세포핵화세포질。결론 FKBP25결실돌변체재HEK 293T、COS7세포중능구표체，위료해FKBP25결실돌변체적공능제공료일정기출。
Objective To investigate the expression and localization of the product of FKBP25 deleted mutants in cells. Methods The full length cDNA 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 immunofluorescence. 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 FKBP25 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.