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Fig. 3 | BMC Microbiology

Fig. 3

From: Production of infectious HCV genotype 1b virus in cell culture using a novel Set of adaptive mutations

Fig. 3

Viral particle production of chimeric sAH-N3.5/TPF1-4B virus. a Organization of full-length HCV constructs. Four sAH-N3.5/TPF1-4B clones were developed by swapping regions of the TPF1-4B genome (from the core to NS2, NS3 to NS5B, the core to E2, and p7 to NS2 regions) with the corresponding region of the sAH-N3.5 genome to create the sAH-N3.5/AgeBsr, /BsrXba, /AgeBbv, and /BbvBsr constructs, respectively. The sAH-N3.5 genome sequences are shown in gray, and the TPF1-4B sequences are shown in white. Huh7 cells were transfected with the full-length RNAs shown on the left, and the amounts of HCV core Ag were determined in the culture medium of cells at 72 h after transfection. b Organization of full-length HCV constructs. Three sAH-N3.5/TPF1-4B clones were generated by swapping the regions NS2, p7 to NS2aa114, and the C-terminal half of NS2 in the TPF1-4B genome with the corresponding regions of the sAH-N3.5 genome to create the sAH-N3.5/XmaBsr, /BbvNsi, and /NsiBsr constructs, respectively. The positions of the three different amino acid variants (Q148R, M170T, and S189L) were confirmed by alignment of the C-terminal half of the NS2 sequences from TPF1-4B and sAH-N3.5 (top row). Three different amino acids sequences were inserted into the TPF1-4B genome to create the TPF1-Q148R, TPF1-M170T, and TPF1-S189L constructs. c In order to compare their ability to produce viral particles, we collected the culture medium of cells at 72 h after transfection and quantified infectivity by FFA. All assays were conducted two independent experiments, and data are represented by an average

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