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Basic Characteristics of Mutations
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Mutation Site
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T563I |
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Mutation Site Sentence
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Although the mutations in E2 and p7 proteins (T563I and N765D) both increased virus infectivity, the patterns within infected cells were different for each, with the E2 mutation (JFH-G m2) exhibiting a dispersed intracellular distribution and p7 (JFH-G m3) showing infection of cells at focal regions (compare panels d and e in Fig. 3C). |
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Mutation Level
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Amino acid level |
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Mutation Type
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Nonsynonymous substitution |
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Gene/Protein/Region
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E2 |
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Standardized Encoding Gene
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E2
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Genotype/Subtype
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- |
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Viral Reference
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-
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Functional Impact and Mechanisms
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Disease
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HCV Infection
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Immune
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- |
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Target Gene
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-
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Clinical and Epidemiological Correlations
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Clinical Information
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- |
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Treatment
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- |
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Location
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- |
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Literature Information
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PMID
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21829654
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Title
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Generation of a cell culture-adapted hepatitis C virus with longer half life at physiological temperature
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Author
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Kim CS,Keum SJ,Jang SK
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Journal
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PloS one
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Journal Info
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2011;6(8):e22808
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Abstract
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BACKGROUND: We previously reported infectious HCV clones that contain the convenient reporters, green fluorescent protein (GFP) and Renilla luciferase (Rluc), in the NS5a-coding sequence. Although these viruses were useful in monitoring viral proliferation and screening of anti-HCV drugs, the infectivity and yield of the viruses were low. METHODOLOGY/PRINCIPAL FINDINGS: In order to obtain a highly efficient HCV cultivation system, we transfected Huh7.5.1 cells [1] with JFH 5a-GFP RNA and then cultivated cells for 20 days. We found a highly infectious HCV clone containing two cell culture-adapted mutations. Two cell culture-adapted mutations which were responsible for the increased viral infectivity were located in E2 and p7 protein coding regions. The viral titer of the variant was approximately 100-fold higher than that of the parental virus. The mutation in the E2 protein increased the viability of virus at 37 degrees C by acquiring prolonged interaction capability with a HCV receptor CD81. The wild-type and p7-mutated virus had a half-life of approximately 2.5 to 3 hours at 37 degrees C. In contrast, the half-life of viruses, which contained E2 mutation singly and combination with the p7 mutation, was 5 to 6 hours at 37 degrees C. The mutation in the p7 protein, either singly or in combination with the E2 mutation, enhanced infectious virus production about 10-50-fold by facilitating an early step of virion production. CONCLUSION/SIGNIFICANCE: The mutation in the E2 protein generated by the culture system increases virion viability at 37 degrees C. The adaptive mutation in the p7 protein facilitates an earlier stage of virus production, such as virus assembly and/or morphogenesis. These reporter-containing HCV viruses harboring adaptive mutations are useful in investigations of the viral life cycle and for developing anti-viral agents against HCV.
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Sequence Data
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-
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