Nucleic acid construct containing full length genome of human hepatitis C virus, recombinant full length virus genome-replicating cells having the nucleic acid construct transferred thereinto and method of producing hepatitis C virus particle
The present invention provides a method for replicating efficiently an RNA containing fulllength HCV genomic sequence and a method for producing HCV virus particles containing fulllength HCV replicon RNA or fulllength HCV genomic RNA by using a cell culture system. Further, the present invention relates to a method for producing hepatitis C virus particles which comprises culturing a cell, into which a replicon RNA comprising a nucleotide sequence comprising a fulllength genomic RNA sequence of hepatitis C virus of the genotype 2a, at least one selectable marker gene and/or at least one reporter gene and at least one IRES sequence or the fulllength genomic RNA of hepatitis C virus of the genotype 2a is introduced, and generating virus particles in the culture medium. Still further the present invention relates also to a hepatitis C vaccine and an antibody against hepatitis C virus particles.
1. A method for producing a cell which replicates an RNA and produces a virus particle, comprising introducing an isolated RNA sequence comprising the nucleotide sequence shown in SEQ ID NO: 12 into a cell selected from the group consisting of a Huh7 cell, a HepG2 cell, a IMY-N9 cell, a HeLa cell and a 293 cell.
2. A method for producing a hepatitis C virus particle, comprising introducing into an isolated RNA sequence comprising the nucleotide sequence shown in SEQ ID NO: 12 into a cell selected from the group consisting of a Huh7 cell, a HepG2 cell, a IMY-N9 cell, a HeLa cell and a 293 cell and culturing the cell to allow the cell to produce a virus particle.
3. A method for producing a virus vector comprising a foreign gene, comprising inserting an isolated RNA sequence encoding a foreign gene into an RNA comprising the nucleotide sequence shown in SEQ ID NO: 12, introducing it into a cell selected from the group consisting of a Huh7 cell, a HepG2 cell, a IMY-N9 cell, a HeLa cell and a 293 cell, and culturing the cell to allow the cell to produce a virus particle.
4. The method of claim 2 , which further comprises obtaining the virus particle from a supernatant of the culture.
5. The method of claim 3 , which further comprises obtaining the virus particle from a supernatant of the culture.