TAILIEUCHUNG - hepatology 2010_part3

Gan - Một cuốn sách giáo khoa lâm sàng hiệu quả lựa chọn của các tế bào thành công transfected và cistron thứ hai có chứa các IRES EMCV chỉ đạo dịch của các protein HCV-cụ thể. | Hepatology - A clinical textbook efficient selection of successfully transfected cells and the second cistron containing the EMCV IRES directs translation of the HCV-specific proteins. Only some Huh7 clones can replicate replicon-specific RNA in titres of approximately 108 positive-stranded RNA copies per microgram total RNA. Moreover all encoded HCV proteins are detected predominantly in the cytoplasm of the transfected Huh7 cells. The development of this replicon is a milestone in HCV research with regard to the investigation of HCV RNA replication and HCV protein analyses. More recently the methodology has been improved in order to achieve significantly higher replication efficiency. Enhancement of HCV RNA replication was achieved by the use of replicons harbouring cell culture-adapted point mutations or deletions within the NS genes Blight 2000 Lohmann 2001 Krieger 2001 . Further development has led to the generation of selectable full-length HCV replicons . genomic replicons that also contain genetic information for the structural proteins Core E1 and E2 Pietschmann 2002 Blight 2002 . This improvement offered the opportunity of investigating the influence of the structural proteins on HCV replication. Thus it has been possible to analyse the intracellular localisation of these proteins. However using this methodology viral assembly and release has not been achieved. Another important milestone was reached when a subgenomic replicon based on the HCV genotype 2a strain JFH-1 was generated Kato 2003 . This viral strain derived from a Japanese subject with fulminant hepatitis C Kato 2001 . The corresponding replicons showed higher RNA replication efficiency than previous replicons. Moreover cell lines distinct from Huh7 such as HepG2 or HeLa were transfected efficiently with transcripts derived from the JFH-1 replicon Date 2004 Kato 2005 . HCV pseudotype virus particles HCVpp . The generation of retroviral pseudotypes bearing HCV E1 and E2 glycoproteins HCVpp .

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