HCV 复制子的研究与应用PPT.ppt

HCV 复制子的研究与应用;HCV研究存在的问题 ;HCV 细胞模型的发展; structural;HCV基因组的异质性 ;HCV复制子研究中的重要问题;Schematic representation of HCV RNAs. The first 21 amino acids of the core coding region (solid box), the gene (Neo, shaded box), the EMCV IRES (EMCV, solid line), H or Con1 indicate H77- or Con1-derived sequences.;细胞的容受性 ;适应性变异的常见位点;Location of cell culture adaptive mutations and increase of G418 transduction efficiency ;Colony-forming abilities of H77 subgenomic RNAs containing mutations in NS3.;Detection of HCV proteins and RNA in Huh-7.5 cells transiently transfected with subgenomic and full-length HCV RNAs.;Effects of cell culture adaptive mutation on infectivity in vivo;Replication of HCV RNAs with and without heterologous elements.;表达NS3蛋白的细胞比例。 FSC-H forward scatter FL1-H fluorescence;HCV复制子研究现状;HCV复制子的应用;HCV复制相关基因结构的研究;The variable region in the 3 NTR enhances RNA replication. ; Determination of the minimal length of the poly(U/UC) tract required for RNA replication. (A) Result of a transient-replication assay using luciferase replicons that carry a homopolymeric uridine tract of 1, 6, 26, or 46 nucleotides. (B) Number of Geneticin-resistant colonies obtained after transfection of selectable replicons with the same modifications in the 3 NTR in panel A. Representative results obtained after transfection of 100 ng of in vitro transcript are shown. (C) Sequence analysis of the poly(U/UC) tracts of replicons isolated from Huh-7 cells that had been transfected with U6 replicons and subjected to selection with Geneticin. ;抗病毒药物的筛选;Antiviral effects of IFN-α , Poly(I)-poly (c) ,and IFN-γa ;92例患者HCV基因分型系谱图;92例患者HCV基因型分布图 ;引物设计图;HCV 1b 型长链 RT- nested PCR及全长基因组产物电泳图