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4 results for “ORF1”
Recombinant Hepatitis E Viruses Harboring Tags in the ORF1 Protein
<p>Hepatitis E virus (HEV) is one of the most common causes of acute hepatitis and jaundice in the world. Current understanding of the molecular virology and pathogenesis of hepatitis E is incomplete, especially due to the limited availability of functional tools. Here, we report the development of tagged HEV genomes as a novel tool to investigate the viral life cycle. A selectable subgenomic HEV replicon was subjected to random 15-nucleotide sequence insertion using transposon-based technology. Viable insertions in the open reading frame 1 (ORF1) protein were selected in a hepatoblastoma cell line. Functional insertion sites were identified downstream of the methyltransferase domain, in the hypervariable region (HVR) and between the helicase and RNA-dependent RNA polymerase domains. HEV genomes harboring a hemagglutinin (HA) epitope tag or a small luciferase (NanoLuc) in the HVR were found to be fully functional and to allow for the production of infectious virus. NanoLuc allowed to quantitatively monitor HEV infection and replication by luciferase assay. HA-tagged replicons and full-length genomes allowed to localize putative sites of HEV RNA replication by the simultaneous detection of viral RNA by fluorescence in situ hybridization and of ORF1 protein by immunofluorescence. Candidate HEV replication complexes were found in cytoplasmic dot-like structures which partially overlap with ORF2 and ORF3 proteins as well as exosomal markers. Hence, tagged HEV genomes yield new insights into the viral life cycle and should allow to further investigate the structure and composition of the viral replication complex.</p>
The unique proteasomal processing of viral ORF1 drives HEV-induced liver fibrosis
GEO Series GSE251924. Homo sapiens. 6 samples. Type: Expression profiling by high throughput sequencing.
Characterization of transcriptome changes induced in 3T3L1 adipocytes by expression of Adenovirus subtype 5 protein E4-ORF1
GEO Series GSE89621. Mus musculus. 6 samples. Type: Expression profiling by high throughput sequencing.
Dataset for Publication "A Modular Hepatitis E Virus Replicon System for Studies on the Role of ORF1-Encoded Polyprotein Domains"
<p>Luciferase Assay readout data of transfection experiments with HEV-based subgenomic reporter replicons. Part of Dissertation "Phylogenetic and Reverse Genetics Studies on Zoonotic Hepatitis E Virus"</p>
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