稳定表达HBx的人肝细胞系7702的建立

doi:10.3969/j.issn.1672-5069.2020.03.005

Abstract

Abstract: Objective The aim of this study was to construct and select an HL-7702 cell line stably expressing HBx in vitro. Methods The HBx gene fragment was amplified by RT-PCR and ligated into pIRES vector. The sequence of pIRES-HBx recombinant plasmid was identified by restriction enzyme digestion and sequencing. The recombinant plasmid was further verified by real-time PCR and Western blot, and the HL-7702 cell stably expressing HBx was selected by G418. Results The correct HBx fragment was amplified by PCR, and the pIRES-HBx plasmid was successfully constructed. The recombinant plasmid overexpressed HBx protein in both HEK 293 cells and HL-7702 cells. The HL-7702 cell stably expressing HBx was further verified by immunofluorescence. Conclusions We Successfully construct a pIRES-HBx expression vector and a stably expressing HBx HL-7702 cells, which might provide us a basic experimental tools for further research.

Key words: HBx, HL-7702 cells, Transfection, In vitro

Pang Lijun, Shi Ying, Guo Xianghua, et al.. Construction and selection of normal human liver 7702 cells stably expressing HBx in vitro[J]. Journal of Practical Hepatology, 2020, 23(3): 320-323.

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Construction and selection of normal human liver 7702 cells stably expressing HBx in vitro

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