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- W2378731423 abstract "Objective To establish a retroviral vector for reversible hepatocyte immortalization.Methods The enhanced green fluorence protein(EGFP) and thymokinase(TK) were amplified and fused.The fusion gene EGFP-TK was cloned to pBABE-puro-lox to construct pBGTKlox,then the immortalized SV40T was connected to the pBGTKlox to establish pBTGTKlox,and finally Cre-ER was connected to pBTGTKlox for establish pCTGTKlox.PT67 cells were co-transfected with pCTGTKlox and pPDF15 and the expression of green fluorescence protein was observed under fluorence microscope.The steadily transfected cells were stained immunohistochemically to detect the expression of SV40 large T antigen.Results A new vector pCTGTKlox containing Cre-ER,SV40T,EGFP-TK and LoxP was constructed and verified by restriction endonuclease digestion.DNA sequencing demonstrated that there was no coding mutation in pCTGTKlox.After co-transfection with pCTGTKlox and pPDF15,PT67 cells emitted green fluorescence under the fluorescence microscope.The nuclei of the transfected cells were positive for immunochemical staining.Conclusion The reversible vector pCTGTKlox is successfully constructed and expressed,thus it provides an ideal vector in the reversible immortalization of hepatocytes and other mammalian cells." @default.
- W2378731423 created "2016-06-24" @default.
- W2378731423 creator A5045392886 @default.
- W2378731423 date "2006-01-01" @default.
- W2378731423 modified "2023-09-23" @default.
- W2378731423 title "Construction,characterization,transfection and expression of vector pCTGTKlox for hepatocyte reversible immortalization" @default.
- W2378731423 hasPublicationYear "2006" @default.
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