Objective: To establish a method for efficient transfer of 1.3-fold HBV/C genome into HepG2 cell line using adenoviral vector system for studying the replication and antigen expression of HBV.
Methods: The 1.3-copy overlength genome of HBV genotype C was constructed and cloned into the shuttle vector pAdTrack. After confirmation of the constructed HBV genome by sequencing, the resultant plasmid linearized by digestion with Pme I was transformed into competent E.coli Adeasier-1 cells. The recombinants of pAdEasy-HBV/C were linearized by digestion with Pac I and transfected into the packaging cells (293 cells) via liposome. HepG2 cells were then infected with a proper quantity of the recombinant adenoviruses. The HBV DNA level and HBeAg and HBsAg titers were detected in the cell medium.
Results: The 1.3-fold overlength HBV/C genome was efficiently transferred into HepG2 cells via the adenoviral vector system, which resulted in initiation of the virus replication and protein expression in the cells using the viral replication mechanism.
Conclusion: The adenovirus vector system (AdEasy) allows convenient and effective transfer of HBV genome into HepG2 cells, and provides a convenient means for screening therapeutic drugs against HBV.
Download full-text PDF |
Source |
|---|
Publication Analysis
Top Keywords
Similar Publications
CRISPR/Cas9 as a New Antiviral Strategy for Treating Hepatitis Viral Infections.
Int J Mol Sci
December 2023
The Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Novosibirsk 630090, Russia.
Hepatitis is an inflammatory liver disease primarily caused by hepatitis A (HAV), B (HBV), C (HCV), D (HDV), and E (HEV) viruses. The chronic forms of hepatitis resulting from HBV and HCV infections can progress to cirrhosis or hepatocellular carcinoma (HCC), while acute hepatitis can lead to acute liver failure, sometimes resulting in fatality. Viral hepatitis was responsible for over 1 million reported deaths annually.
View Article and Find Full Text PDFMolecular epidemiology of hepatitis B among Indigenous Australians in Queensland and the Torres Strait Islands.
Intern Med J
January 2024
Gallipoli Medical Research Institute, Greenslopes Private Hospital, Brisbane, Queensland, Australia.
Background: Chronic hepatitis B virus (HBV) infection is a major health problem for all Indigenous Australians. Post-2000, Hepatitis B surface antigen prevalence has decreased, although remaining four times higher among Indigenous compared with non-Indigenous people.
Aims: This study aimed to characterise the HBV from Indigenous populations in Queensland and the Torres Strait Islands.
[Distributive characteristics of HBV DNA CpG islands in HBsAg positive mothers and its relationship with intrauterine transmission].
Zhonghua Liu Xing Bing Xue Za Zhi
May 2022
Department of Epidemiology/Center of Clinical Epidemiology and Evidence-Based Medicine, Shanxi Medical University, Taiyuan 030001, China.
To investigate the type, length, and CG loci of HBV DNA CpG islands in HBsAg positive maternal C genotype and its relationship with intrauterine HBV transmission, so as to provide a new perspective for the study of intrauterine transmission of HBV. From June 2011 to July 2013, HBsAg-positive mothers and their newborns who delivered in the obstetrics and gynecology department of the Third People's Hospital of Taiyuan were collected. Epidemiological data were collected through face-to-face questionnaires and electronic medical records.
View Article and Find Full Text PDFDating the origin and dispersal of global hepatitis B virus genotype C in humans.
Drug Discov Ther
May 2022
School of Public Health, Shenzhen University Health Science Center, Shenzhen, China.
Hepatitis B virus genotype C (HBV/C) is one of the most prevalent HBV strains worldwide, especially in the Western Pacific and the South-East Asia. However, the origin and evolutionary timescale of HBV/C remains largely unresolved. We analyzed the evolutionary rate and molecular clock phylogeny of 101 full-genome HBV/C sequences sampled globally using a Bayesian Markov Chain Monte Carlo (MCMC) approach.
View Article and Find Full Text PDF[Role of the liver-enriched transcription factor binding site mutation in the C promoter region of hepatitis B virus genome for HBx-enhanced hepatitis B virus replication].
Zhonghua Gan Zang Bing Za Zhi
April 2021
Center of Infectious Diseases, West China Hospital of Sichuan University, Chengdu 610041, China.
To construct a recombinant HBV replication-type plasmid with liver-enriched transcription factor binding site mutation at proximal of HBV C promoter in order to elucidate the role of HBx-enhanced HBV replication. Site-directed mutagenesis technology was used to construct a recombinant plasmid with liver-enriched transcription factor binding site mutation at proximal of HBV C promoter on the basis of wild-type HBV replicating plasmid and HBV replicating plasmid lacking HBx expression. Subsequently, plasmid transfection was carried out in HBV liver cancer cell replication model and mouse replication model, and HBV replication intermediates of cells and mouse liver tissue were extracted for detection.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!