Expression of hepatitis B virus surface and e antigen genes cloned in bovine papillomavirus vectors.
Sequences of the hepatitis B virus (HBV) genome coding for the surface antigen (HBsAg), but lacking the regulatory (preS) sequences, were cloned into a bovine papillomavirus (BPV) vector consisting of the transforming 69% of the BPV genome (BPV 69T), and transformed into mouse c127 cells. Clones carrying HBV and BPV sequences in the same transcriptional orientation did not produce immunologically active HBsAg, while those with the opposite orientation produced and secreted HBsAg. RNA species of an HBsAg producer were 3400, 9600, 11000 and 18000 nucleotides long and hybridized with both HBV and BPV probes. Mouse cells (c127) transformed with the whole BPV genome (BPV-1) carrying both the HBsAg-coding sequences and the regulatory preS sequences of HBV DNA were stable and produced and secreted HBsAg 22 nm particles. These yielded 11500 and 12500 nucleotide RNA transcripts which also hybridized with both BPV and HBV DNA probes. BPV-1 carrying whole HBV DNA monomer or dimer genomes yielded transformants which initially produced HBsAg as well as HBV e antigen (HBeAg), but these were not stable. The hybrid genomes, with the exception of those carrying the HBV dimers, existed as multicopy plasmids (50-100 copies per cell) and often acquired new sequences.
D000818:Animals; D002417:Cattle; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000235:genetics; D005786:Gene Expression Regulation; D005814:Genes, Viral; D005822:Genetic Vectors; D006511:Hepatitis B Antigens / Q000235:genetics*; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D006513:Hepatitis B e Antigens / Q000235:genetics*; D027383:Papillomaviridae / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics
D004274:DNA, Recombinant; D004279:DNA, Viral; D006511:Hepatitis B Antigens; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D012367:RNA, Viral