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Objective: To and amplification hRev7 green fluorescent plasmid transfected N2A cells and neuronal cells , and to investigate the mechanism of neuronal apoptosis hRev7 . Methods: Preparation of competent cells , plasmids through bacterial inoculation , shake bacteria amplification , plasmids were extracted strip seen in the fluorescence camera system , and using a spectrophotometer to test its purity , built on the green fluorescence was amplified carrier in PEGFP-1 hRev7 cultured N2A cells transiently transfected N2A cells Lipofectamin2000 , Annexin V-APC labeled apoptotic cells , flow cytometry of hRev7 on apoptosis . Results: (1 ) via the bacterial inoculation , preparation of competent cells , plasmids , amplified rocking bacteria , extracted to a higher purity of the plasmid . (2 ) were transiently transfected N2A cells detected by flow cytometry detector containing hRev7 with green fluorescent cells transfected with pEGFP-N1-hRev7 apoptosis rate was 14.8 , and transfected with pEGFP-N1 cells The apoptosis rate of 6.5 , the statistical analysis , the difference was significant ( p lt; 0.05 ) does not reject the null hypothesis , that significant apoptosis rate of the experimental group and the control group, the apoptosis rate differences . The data results show that compared with the control empty plasmid group hRev7 promote apoptosis . Conclusion : Amplification the plasmid of PEGFP-hRev7 , by flow cytometry showed that transient transfection hRev7, N2A cells can promote apoptosis , and laid the foundation for further research Down syndrome and hRev7 of mechanism of apoptosis of neurons .
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