Construction of an immortalized rat astrocyte strain with inducible enkephalin expression
- VernacularTitle:强力霉素调控表达脑啡肽的永生化大鼠星形胶质细胞株的构建
- Author:
Ying XU
;
Yu-Ke TIAN
;
Xue-Bi TIAN
;
- Publication Type:Journal Article
- Keywords:
Enkephalins;
Doxycycline;
Gene expression regulation;
Astrocytes
- From:
Chinese Journal of Anesthesiology
1994;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct an immortalized rat astrocyte strain expressing enkephalin which can be regulated by doxycycline.Methods pRevTet-On and recombinant plasmid pRevTRE/hPPE were transfected into packaging cell PT67 respectively by standard lipofectamine methods.The supernatant containing RevTet-On virus was used to infect immortalized rat astrocyte strain(IAST)and RevTRE/hPPE virus was used to infect the IAST/Tet-On cell.Immortalized rat astrocyte strain that stably expressed Tet-regulated enkephalin was established. hPPE gene expression level of IAST/Tet-On/hPPE cell strain was detected by real time-PCR,immuno- cytochemistry and radio-immunoassay.Results Real time-PCR,immuno-cytochemistry and radio-immunoassay confirmed the level of enkephalin expression was regulated by doxycycline in a dose-dependent manner.Conclusion An immortalized rat astrocyte strain which secrets enkephalin as controlled by doxycycline has been constructed successfully.
