EFFICIENT CELL CULTURE SYSTEM FOR HEPATITIS C VIRUS GENOTYPE 4a
AssigneeHvidovre Hospital
InventorsMartin Schou Pedersen, Jens Bukh, Long Van Pham, Santseharay Ramirez Almeida, Ulrik Fahnøe, Carlota Fernandez-Antunez, Daryl Grant Humes, Kristian Schønning
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Abstract
The present invention relates to nucleic acid sequences that encode hepatitis C viruses (HCV) of genotype da that are useful in the fundamental research of HCV as well as in the search of antivirals and vaccines against HCV. In particular, the present invention relates to nucleic acid sequences that comprise HCV, which are capable of expressing the virus when transfected into cells and are capable of replication or infectivity in cultured cells.
Claims19
An isolated nucleic acid molecule which encodes human hepatitis C virus (HCV) or a HCV replication system of genotype 4a, strain ED43, wherein the said molecule comprises a nucleic acid sequence encoding an amino acid sequence with a sequence identity of at least 99% to SEQ ID NO:
- 1 or a fragment hereof encoding the said HCV replication system corresponding to NS3-NS5B between amino acids 1027-3008 of SEQ ID NO:
- 1 or SEQ ID NO:
- 2, or an amino acid sequence with a sequence identity of at least 99% to the amino acids 1027-3008 of SEQ ID NO:
- 1 or SEQ ID NO:
- 2, and wherein the amino acid sequence or the fragment hereof
- comprises the following adaptive mutations:
- 11291V, S1465G, A1672S, A1786V, T1822A, T1865A, S1870N, D2413G, D2545E, D2675E, K2797R, E2806T, A2916V, D2976G, Y2978F, M2981V, L2991R and C2992Y according to SEQ ID NO:
- 1;
- optionally, further comprises the following adaptive mutation T827A according to SEQ ID NO:
- 1;
- and
- one of the following groups of additional adaptive mutations:
- a) Q2931R according to SEQ ID NO:
- 1;
- b) V271G, C458R, Y848C, L1466M, F1572L, G1909A, A2257T, T2329A, K2597N, V2793A, Q2931R and S2982P according to SEQ ID NO:
- 1;
- or
- c) V271G, C458R, Y848C, L1466M, F1572L, G1909A, A1973T, A2257T, T2329A, K2597N, V2793A, and S2982P according to SEQ ID NO:
- 1.
2-25. (canceled)
A method for screening an anti-hepatitis C virus substance, comprising
- a) culturing a cell comprising:
- a nucleic acid molecule which encodes human hepatitis C virus (HCV) or a HCV replication system of genotype 4a, strain ED43, wherein the said molecule comprises a nucleic acid sequence encoding an amino acid sequence with a sequence identity of at least 99% to SEQ ID NO:
- 1 or a fragment hereof encoding the said HCV replication system corresponding to NS3-NS5B between amino acids 1027-3008 of SEQ ID NO:
- 1 or SEQ ID NO:
- 2, or an amino acid sequence with a sequence identity of at least 99% to the amino acids 1027-3008 of SEQ ID NO:
- 1 or SEQ ID NO:
- 2, and wherein the amino acid sequence or the fragment hereof
- comprises the following adaptive mutations:
- 11291V, S1465G, A1672S, A1786V, T1822A, T1865A, S1870N, D2413G, D2545E, D2675E, K2797R, E2806T, A2916V, D2976G, Y2978F, M2981V, L2991R and C2992Y according to SEQ ID NO:
- 1;
- optionally, further comprises the following adaptive mutation T827A according to SEQ ID NO:
- 1;
- and
- one of the following groups of additional adaptive mutations:
- d) Q2931R according to SEQ ID NO:
- 1;
- e) V271G, C458R, Y848C, L1466M, F1572L, G1909A, A2257T, T2329A, K2597N, V2793A, Q2931R and S2982P according to SEQ ID NO:
- 1;
- or V271G, C458R, Y848C, L1466M, F1572L, G1909A, A1973T, A2257T, T2329A, K2597N, V2793A, and S2982P according to SEQ ID NO:
- 1
- b) detecting the replicating RNA or the virus particles in the resulting culture.
Dependencies are inferred from the claim text.