Dataset Information

DNA Engineering and Hepatitis B Virus Replication.

ABSTRACT: Recombinant DNA technology is a vital method in human hepatitis B virus (HBV), producing reporter viruses or vectors for gene transferring. Researchers have engineered several genes into the HBV genome for different purposes; however, a systematic analysis of recombinant strategy is lacking. Here, using a 500-bp deletion strategy, we scanned the HBV genome and identified two regions, region I (from nt 2,118 to 2,814) and region II (from nt 99 to 1,198), suitable for engineering. Ten exogenous genes, including puromycin N-acetyl transferase gene (Pac), blasticidin S deaminase gene (BSD), Neomycin-resistance gene (Neo), Gaussia luciferase (Gluc), NanoLuc (Nluc), copGFP, mCherry, UnaG, eGFP, and tTA1, were inserted into these two regions and fused into the open reading frames of hepatitis B core protein (HBC) and hepatitis B surface protein (HBS) via T2A peptide. Recombination of 9 of the 10 genes at region 99-1198 and 5 of the 10 genes at region 2118-2814 supported the formation of relaxed circular (RC) DNA. HBV DNA and HBV RNA assays implied that exogenous genes potentially abrogate RC DNA by inducing the formation of adverse secondary structures. This hypothesis was supported because sequence optimization of the UnaG gene based on HBC sequence rescued RC DNA formation. Findings from this study provide an informative basis and a valuable method for further constructing and optimizing recombinant HBV and imply that DNA sequence might be intrinsically a potential source of selective pressure in the evolution of HBV.

SUBMITTER: Gan CY

PROVIDER: S-EPMC8632529 | biostudies-literature |

REPOSITORIES: biostudies-literature

ACCESS DATA

Similar Datasets

Project description:Purpose: Worldwide, over 350 million people are chronically infected with the hepatitis B virus (HBV) and are at increased risk of developing progressive hepatic diseases (hepatitis, cirrhosis or liver cancer). Still, the metabolic processes operational during the distinct clinical phases of a chronic HBV infection to liver disease remains unexplored. Thus, the aim of our study was to determine the serum metabolic characteristics of hepatic disease patients with chronic hepatitis B in active and inactive stage and study the influence of active and inactive stage of the chronic hepatitis B virus on the development of liver cancer by untargeted metabolomics. Method: To investigate this, we conducted an untargeted metabolomics approach to determine the metabolic characteristics of 199 human serum samples from hepatic disease patients in active and inactive chronic HBV via UHPLC-Q-TOF/MS, and patients with HBV in active or inactive stage were grouped based on their serum levels of HBV DNA and HBV envelope antigen (HBeAg). Principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and volcano plot were used for multivariate analysis. Pathway analysis were conducted according to p-values from pathway enrichment analysis and pathway impact values from pathway topology analysis. In addition, commercial databases were used to identify metabolites from significantly altered metabolic pathways. All samples were analyzed for liver function tests indicated the levels of certain enzymes. Results: Our data provide insight into the metabolic dysregulation experienced during active or inactive stage of the HBV on the development of liver disease, and analysis of serum metabolomic profiles indicated that most of the altered metabolic pathways for chronic hepatitis and cirrhosis patients with active or inactive stage of the HBV were associated with D-Glutamine and D-glutamate metabolism, indicating a strong link to HBV replication (p < 0.05). In addition, liver function analysis showed the enzymes gamma-glutamyl transferase (GGT) were increased concomitant to the progression of hepatitis to cirrhosis induced by chronic HBV (p < 0.05).

Dataset Information

DNA Engineering and Hepatitis B Virus Replication.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets