Abstract: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.
