Characterization and Mapping of Viral and Putative Viral-Cellular Transcripts in a Hepatitis B Virus Infected Human Hepatoma Cell Line and in Chimpanzee Carrier Liver
Since attempts to propagate hepatitis B virus (HBV) in tissue culture have generally been unsuccessful, recent studies on the mechanism for viral morphogenesis have focused on expression of the HBV genome in HBV infected human hepatoma cell lines (1,2,3,4) and in vitro transfected rodent cells (5). The human hepatoma cell lines PLC/PRF/5 and HEP-3B synthesize and secrete 22-nanometer particles with immunochemical characteristics of HBsAg and contain HBV DNA exclusively in integrated form (1,2,3,6,7). Initial reports of viral RNAs expressed in PLC/PRF/5 cells have given conflicting results. Marion et al. (1) demonstrated sequences hybridizing with the whole genome of HBV by solution hybridization, whereas Edman et al. (3) found only one transcript of 2000 nucleotides, which hybridized to the S region (HBsAg). However, in L cells transformed with cloned HBV DNA and in PLC/PRF/5 cells, Pourcel et al. (8) recently detected a 2.3 kb HBV-specific poly (A)+ RNA. This RNA hybridized to the pre-S region, to the S region, and to a 1100 bp region downstream from the S gene coding sequence; and it was proposed that the sequence 5′-TATATAA starting at position 2776 could be related to the S gene promoter (8).
KeywordsCellulose Phenol EDTA DMSO Glycine
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