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A Competent Hepatocyte Model Examining Hepatitis B Virus Entry through Sodium Taurocholate Cotransporting Polypeptide as a Therapeutic Target
Published on: May 10, 2022
[The influences of Tet system on activity and tissue specificity of HBV Cp.]
Jun-Chuan Lei1, Wei-Dong Gong, Ya Zhao
1Department of Etiology, School of Basic Medicine, Xi'an 710032, China.
Aim:
Analyze the effect of Tetracycline-controlled system on Hepatitis B virus core promoter (Cp) activity and tissue specificity.
Methods:
The 7 Teto sequence was amplified from plasmid pTL-8 using polymerase chain reaction (PCR) and cloned into T vector pMD19-T Simple. After sequencing, the 7 teto sequence was subcloned into upstream of Cp in pGL3-Basic/Cp. In order to observe the effect of Tetracycline-controlled system on Cp activity and tissue specificity, the plasmid pTL-8 which luciferase encoding sequence was excised and pGL3-Basic/7t/Cp were cotransfected into different tissue-derived cell lines including HepG2, Hela, COS-7, MDA-MB-231 and HT-29. The dual luciferase activities were determined by Dual Luciferase Report (DLR) assay system.
Results:
Have successfully constructed plasmid pGL3-Basic/7t/Cp. The transcriptional activity of Cp was increased significantly after the teto sequence was cloned upstream of Cp.
Conclusion:
The transcriptional activity of Cp is augmented significantly by Tetracycline-controlled system. But the tissue specificity of Cp lost at the same time.
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