[PDF] Top 20 Quantitative Analysis of the Hepatitis C Virus Replication Complex

... (B) Analysis of in vitro replicase activity in total lysates (TL) and different subcellular fractions of replicon cells (left half) and naive Huh-7 cells (right ...hybridization analysis with the same ... See full document

... RNA replication occurs primarily on the outer DMV membrane facing the cytoplasm as proposed for picornavi- ruses (57, 58) or whether it takes place mainly in the DMV lumen as suggested for coronaviruses (51, 52) ... See full document

... JFH-1 replication to uptake and secrete infectious HCV JFH-1 ...gradient analysis and RT-QPCR analysis of HCV RNA isolated from the mouse culture supernatants supports the conclusion that very few, ... See full document

... Protein expression and purification. A construct of the wild-type HCV polymerase genotype 2a JFH-1 isolate was designed with the C-terminal 21 amino acids removed and replaced with a noncleavable hexahistidine ... See full document

... ABSTRACT Hepatitis C virus (HCV) infection has been shown to regulate microRNA 130a (miR-130a) in patient biopsy specimens and in cultured ...and hepatitis B virus (HBV) ...HCV ... See full document

... RNA-protein interaction analysis. For the RNA pulldown assay, we adapted a protocol used previously to identify cellular proteins interacting with the HCV 3=UTR (12). Briefly, a biotin-labeled DNA oligonucleotide ... See full document

... hybridization analysis was carried out to determine the composition of ...labeled replication products were eluted from an agarose gel and used as a probe for hybridization in a Northern blot which included ... See full document

... 3. Replication of a genotype 2a strain, JFH1, was less sensitive to ...RT-PCR analysis, and the relative amounts are ...immunoblot analysis. (C) HCV RNA was quantified and plotted as described ... See full document

... for replication in transient repli- cation assays, and NS5A expression was examined by Western blot analysis following vaccinia virus-T7 polymerase-mediated (VV- T7) expression of the ... See full document

... of replication, but the effects we observe are small, and it is therefore unlikely that serine 222 is a major regulator of RNA ...on virus production, suggesting that different serine phosphory- lation ... See full document

... and replication efficiency in cells with different CD81 expression levels, two cell populations, CD81-high (CD81-H) and CD81- low 1 (CD81-L1), containing 90% and 10% CD81-positive cells, respectively, were first ... See full document

... for quantitative RT-PCR determination of PLA2G4C mRNA and HCV RNA levels after 96 ...blot analysis of PLA2G4C, HCV NS3, and ␤ -actin at 96 ...a quantitative RT-PCR determination of HCV RNA levels (E) ... See full document

... pseudotype virus (25) for 48 h before plaque ...VSV replication, since the VSV titer increases in the presence of ISG56 ...pseudotype virus entry ...HCV replication was measured after 48 h of ... See full document

... qRT-PCR analysis of selected genes revealed an early response of transcrip- tional reprogramming to low oxygen ...(FACS) analysis (see Table S3 in the supplemental material) also supported an acceler- ation ... See full document

... This analysis was performed by inoculating culture medium that contained extracellular virions and cell extracts that contained intracellular virions, respectively, into naive ...RNA replication was ... See full document

... for replication (26), we first tested the tolerance for reporter expression in place of deleted ...and C termini of the reporter, based on the hypothesis that processing of the re- porter from nsp1 and nsp3 ... See full document

... miR-122:HCV complex should not be affected by this ...the replication defect of HCV site 2, although the replication efficiency was only one-fifth that of the WT ...HCV replication, although ... See full document

... RNA replication. For instance, the 3⬘ NTR of GB virus B, a virus that is very closely related to HCV, also has a tripartite structure com- posed of a variable region, a poly(U) tract of 9 to 30 ... See full document

... during replication complex formation and viral RNA synthesis; however, the specific functions of many nsp during virus replication remain ...basic replication and pathogenesis ... See full document

... several virus species has allowed to elucidate functions of viral gene ...RNA replication, i.e., the viral replication complex, have been pinpointed in such experiments for poliovirus, ... See full document