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- aggregation classification "A1".
- aggregation creator B183688.
- aggregation creator B183689.
- aggregation creator B183690.
- aggregation creator B183691.
- aggregation creator B183692.
- aggregation creator B183693.
- aggregation creator B183694.
- aggregation creator B183695.
- aggregation creator person.
- aggregation creator person.
- aggregation date "2011".
- aggregation format "application/pdf".
- aggregation hasFormat 2066101.bibtex.
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- aggregation isPartOf urn:issn:0022-538X.
- aggregation language "eng".
- aggregation rights "I have transferred the copyright for this publication to the publisher".
- aggregation subject "Biology and Life Sciences".
- aggregation title "Development and application of hepatitis C reporter viruses with genotype 1 to 7 core-nonstructural protein 2 (NS2) expressing fluorescent proteins or luciferase in modified JFH1 NS5A".
- aggregation abstract "To facilitate genotype-specific high-throughput studies of hepatitis C virus (HCV), we have developed reporter viruses using JFH1-based recombinants expressing core-nonstructural protein 2 (NS2) of genotype 1 to 7 prototype isolates. We introduced enhanced green fluorescent protein (EGFP) into NS5A domain III of the genotype 2a virus J6/JFH1 [2a(J6)]. During Huh7.5 cell culture adaptation, 2a(J6)-EGFP acquired a 40-amino-acid (aa) (Delta 40) or 25-aa (Delta 25) deletion in NS5A domain II, rescuing the impairment of viral assembly caused by the EGFP insertion. Delta 40 conferred efficient growth characteristics to 2a(J6) tagged with EGFP, DsRed-Express2, mCherry, or Renilla luciferase (RLuc), yielding peak supernatant infectivity titers of 4 to 5 log(10) focus-forming units (FFU)/ml. 2a(J6) with Delta 40 or Delta 25 was fully viable in Huh7.5 cells. In human liver chimeric mice, 2a(J6)-EGFP Delta 40 acquired various deletions in EGFP, while 2a(J6)Delta 40 did not show an impaired viability. We further developed panels of JFH1-based genotype 1 to 7 core-NS2 recombinants expressing EGFP- or RLuc-NS5A Delta 40 fusion proteins. In cell culture, the different EGFP recombinants showed growth characteristics comparable to those of the nontagged recombinants, with peak infectivity titers of 4 to 5 log(10) FFU/ml. RLuc recombinants showed slightly less efficient growth characteristics, with peak infectivity titers up to 10-fold lower. Overall, the EGFP and RLuc recombinants were genetically stable after one viral passage. The usefulness of these reporter viruses for high-throughput fluorescence- and luminescence-based studies of HCV-receptor interactions and serum-neutralizing antibodies was demonstrated. Finally, using RLuc viruses, we showed that the genotype-specific core-NS2 sequence did not influence the response to alfa-2b interferon (IFN-alfa-2b) and that genotype 1 to 7 viruses all responded to treatment with p7 ion channel inhibitors.".
- aggregation authorList BK440074.
- aggregation endPage "8928".
- aggregation issue "17".
- aggregation startPage "8913".
- aggregation volume "85".
- aggregation aggregates 2082639.
- aggregation isDescribedBy 2066101.
- aggregation similarTo JVI.00049-11.
- aggregation similarTo LU-2066101.