文章摘要
王变荣,张颖冬,石静萍.稳定表达AT1R基因且分泌Aβ的HEK293细胞株构建[J].南京医科大学学报,2010,(12):1703~1707
稳定表达AT1R基因且分泌Aβ的HEK293细胞株构建
Establishment of a stable AT1R-expressed and Aβ secretion HEK293 cell line
  
DOI:10.7655
中文关键词: 血管紧张素Ⅱ1型受体  APP-PS1-HEK293  基因转染  基因表达
英文关键词: AT1R  APP-PS1-HEK293  gene transfection  gene expression
基金项目:南京市医学重点科技发展项目资助(ZKX08035)
作者单位
王变荣 南京大学医学院神经病学系,江苏 南京 210093 
张颖冬 南京医科大学附属脑科医院神经内科,江苏 南京 210029 
石静萍 南京医科大学附属脑科医院神经内科,江苏 南京 210029 
摘要点击次数: 1395
全文下载次数: 111
中文摘要:
      目的: 用人血管紧张素Ⅱ1型受体(AT1R)基因转染表达人Aβ前体蛋白(APP)及早老素蛋白1(PS1)的人胚肾293(APP-PS1-HEK293)细胞,建立稳定转染细胞系?方法:利用真核表达载体pcDNA3.1-AT1R以阳离子脂质体法转染APP-PS1-HEK293细胞,pcDNA3.1-EGFP为阴性对照,通过G418(600 μg/ml)选择培养,建立稳定转染细胞系,RT-PCR?细胞免疫荧光及Western blot检测转染的AT1R的表达?结果:pcDNA3.1-AT1R稳定转入APP-PS1-HEK293细胞,稳定成功表达目的基因?结论:稳定转染细胞系的建立和基因表达为进一步研究AT1R对Aβ分泌的影响及其在阿尔茨海默病发病机制探讨中提供了良好实验基础?
英文摘要:
      Objective: To transfect human angiotensin Ⅱtype 1 receptor(AT1R)gene to human embryonic kidney(HEK)293 cells coexpressing human amyloid-beta protein precursor(APP)and presenilin-1(PS1)gene(APP-PS1-HEK293 cells),select high AT1R expression clone,and verify. Methods:Using pcDNA3.1-AT1R to transfect APP-PS1-HEK293 cells,pcDNA3.1-EGFP as control,selecting positive clones by G418(600 μg/mL),amplifying,then verifying them by RT-PCR,Immunofluorescence and Western blot. Results:RT-PCR,Immunofluorescence and Western blotting indicated that APP-PS1-HEK293 transfected by AT1R gene highly and stably expressed AT1R. Conclusion:The establishment of the stably transfected cell line expressing the target gene provides a solid experimental foundation for further studies on the role of the AT1R gene in the pathogenesis of Alzheimer’s disease.
查看全文   查看/发表评论  下载PDF阅读器
关闭