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Investigating the function of Akt by tet-off inducible expression system
Authors:Qiwei Zhai  Hongbin Ji  Zhongcheng Zheng  Lanying Sun  Xinyuan Liu
Institution:Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031 ,China
Abstract: A tet-off inducible cell line named BBT derived from BA/F3b cell line was constructed and the effect of this inducible expression system was significant when detected by tet-off responded luciferase reporter gene assay. Then tet-off responded Akt expression plasmid was transfected into BBT cells, and the stable cell lines were screened. The result of Northern blot showed that the expression of akt was significantly inducible. The clone with the best inducible effect was selected and named BBA for investigating the function of Akt. We found that Akt could significantly inhibit zinc-induced decrease of cell viability when assayed by MTT method. And the flow cytometric analysis showed that Akt could markedly repress zinc-induced apoptosis.
Keywords:inducible expression system  Akt  apoptosis
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