RESUMEN
Hepatitis delta virus (HDV) is a replication-defective etiological agent of hepatitis that requires hepatitis B virus (HBV) as a helper. A complementary DNA (cDNA) fragment of the RNA genome of HDV was cloned into the plasmid vector pBR322, and the primary nucleotide sequence and predicted protein products of the cDNA fragment were determined. This cloned cDNA fragment has been used as a sensitive radioactive probe for the detection of HDV RNA in the serum of patients with either acute or chronic HDV infections.
Asunto(s)
Hepatitis D/diagnóstico , Virus de la Hepatitis Delta/genética , ARN Viral/genética , Secuencia de Bases , Clonación Molecular , Hepatitis D/microbiología , Humanos , Hibridación de Ácido Nucleico , Pan troglodytesRESUMEN
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.
Asunto(s)
Antígenos de la Hepatitis B/genética , Antígenos de Superficie de la Hepatitis B/genética , Antígenos e de la Hepatitis B/genética , Animales , Bovinos , Clonación Molecular , ADN Recombinante , ADN Viral/genética , Regulación de la Expresión Génica , Genes Virales , Vectores Genéticos , Papillomaviridae/genética , ARN Viral/genéticaRESUMEN
We have determined the sequences of the ori region DNA of several phage lambda mutants and hybrids, which shed light on the mechanism of DNA replication in the lambdoid phages. These include the heterologous substitution hybrids lambda rep82:lambda and lambda rep80:lambda, a pseudorevertant of the ori-r93 mutant lambda r93hot5, and the insertion mutant lambda pk35. The ori regions of the three lambdoid phages, lambda, phi 80 and 82, all have repeated sequences, termed iterons, and A . T-rich zones. We note that a similar arrangement of DNA is also found in several other prokaryotic origins of replication. lambda and phi 80 have four iterons, and 82 has five. The origin of lambda r93hot5 is unusual in that contains only three iterons, yet the phage grows normally. Analysis of this mutant indicates that the spacing of iterons is crucial to ori function, whereas their number is not. This argues against the cloverleaf model for lambda ori structure (Hobom et al., 1979). In lambda pk35 the drug resistance element Tn903 is inserted into the "inceptor" (ice) site, proposed to be crucial for lambda replication initiation (Hobom et al., 1979); yet this phage grows normally.
Asunto(s)
Bacteriófago lambda/genética , Replicación del ADN , Secuencia de Bases , Genes Reguladores , Genes Virales , Conformación Proteica , Secuencias Repetitivas de Ácidos Nucleicos , Replicón , Proteínas Virales/genéticaRESUMEN
We describe the characterization of 34 hybrid lambda bacteriophages carrying EcoRI fragments obtained from DNA of defective interfering particles of the Patton strain of Herpes simplex virus type 1 (HSV-1). All cloned fragments contained S region terminal repeat sequences (TRs) fused to unique HSV-1 DNA. Several fragments contained deletions and rearrangements not described previously for DNA of HSV-1 defective interfering particles. A model describing the generation of defective interfering DNA based on recombination events involving the terminal "a" sequence as presented.
Asunto(s)
Bacteriófago lambda/genética , ADN Recombinante/análisis , ADN Viral/análisis , Simplexvirus/genética , Secuencia de Bases , Mapeo Cromosómico , Replicación del ADN , Enzimas de Restricción del ADN/metabolismo , Secuencias Repetitivas de Ácidos NucleicosRESUMEN
Biochemical and electron microscopic data indicate that the human hepatitis delta viral agent contains a covalently closed circular and single-stranded RNA genome that has certain similarities with viroid-like agents from plants. The sequence of the viral genome (1,678 nucleotides) has been determined and an open reading frame within the complementary strand has been shown to encode an antigen that binds specifically to antisera from patients with chronic hepatitis delta viral infections.