Localization on the viral genome and nucleotide sequence of the gene coding for the two major polypeptides of the Hepatitis B surface antigen (HBs Ag)

Charnay, Patrick; Mandart, Elisabeth; Hampe, A; Fitoussi, Françoise; Tiollais, Pierre; Galibert, Francis

doi:10.1093/nar/7.2.335

Cited by 74 publications

(37 citation statements)

References 29 publications

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“…Nucleic acid homologies between the cloned HBV and WHV DNAs were detected by Southern gel hybridization (3). Cloning of the complete HBV DNA genome by using XgtWES-XB and subsequent studies using this clone have been reported (4)(5)(6)(7).…”

Section: Recently Summers Et Al (1) Demonstrated That the Human Hepamentioning

confidence: 99%

Isolation, characterization, and comparison of recombinant DNAs derived from genomes of human hepatitis B virus and woodchuck hepatitis virus.

Cummings¹,

Browne²,

Salser³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

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The human hepatitis B virus (HBV) and the woodchuck hepatitis virus (WHV) are closely related by several criteria and belong to the same class of DNA viruses. The DNA genomes from these viruses are difficult to obtain in quantities required for biochemical analysis. We have, therefore, cloned these two DNAs in the vector XgtWES and subcloned into the kanamycin resistance plasmid pAOl. Comparison of the recombinant DNAs with authentic viral DNAs by specific hybridization, size, and restriction enzyme analysis suggests that the recombinants contain the complete genome of each virus. The nominal size of the cloned HBV genome was 3150 base pairs, compared to 3200 base pairs for the cloned WHV genome. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

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Section: Recently Summers Et Al (1) Demonstrated That the Human Hepamentioning

confidence: 99%

Isolation, characterization, and comparison of recombinant DNAs derived from genomes of human hepatitis B virus and woodchuck hepatitis virus.

Cummings¹,

Browne²,

Salser³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

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“…The antigenicity and immunogenicity of this protein (designated S-protein) depends on the maintenance of disulphide bonds (Vyas et al, 1972;Sukeno et al, 1972;Dreesman et al, 1973). The open reading frame on HBV DNA coding for S protein (Charnay et al, 1979;Peterson et al, 1977) has the capacity to code for a protein consisting of 389 to 400 amino acids (depending on the antigenic subtype of HBV). The DNA sequence corresponding to this reading flame and preceding the gene for S-protein (i.e.…”

Section: Introductionmentioning

confidence: 99%

Detection of Antiviral Antibodies with Predetermined Specificity Using Synthetic Peptide-beta-Lactamase Conjugates: Application to Antibodies Specific for the preS Region of the Hepatitis B Virus Envelope Proteins

Neurath¹,

Kent²,

Strick³

1986

Journal of General Virology

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SUMMARYAmino acid sequences coded for by the preS region of the hepatitis B virus (HBV) envelope gene are present both in HBV and in subviral hepatitis B surface antigen (HBsAg) particles. Consequently, anti-preS-specific antibodies are elicited during the course of HBV infection. Such antibodies are virus-neutralizing. Therefore, it is important to determine whether or not vaccination with HBsAg also induces an antipreS-specific immune response. We describe here an enzyme-linked immunosorbent assay applicable for the screening of sera from vaccinated individuals for anti-preS antibodies. IgG from serum specimens was adsorbed to staphylococcal Protein A on a superparamagnetic support and subsequently mixed with a synthetic peptide analogue [preS(120-145)] covalently linked to fl-lactamase. The presence of anti-preS in serum specimens resulted in binding of the conjugated fl-lactamase to the magnetic support. The adsorbed enzyme was quantified colorimetrically.

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“…Recently, information on the genetic organization of the virus was obtained from the nucleotide sequence of the genome. Genes coding for surface and capsid antigens and perhaps for DNA polymerase are located on the long strand of the genome (4)(5)(6)(7). However, nothing is known concerning virus gene expression and its regulation.…”

mentioning

confidence: 99%

Excretion of hepatitis B surface antigen particles from mouse cells transformed with cloned viral DNA.

Dubois¹,

Pourcel²,

Rousset³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

211

123

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A plasmid containing two cloned he atitis B virus genomes in a tandem head-to-tail arrangement gas been introduced into mouse fibroblasts by using cotransformation with the cloned herpes simplex virus thymidine kinase gene. Several copies of the plasmid were integrated into high molecular weight cellular DNA. The original tandem structure of the hepatitis B virus DNA was conserved. Hepatitis B surface antigen was synthesized by all the 15 clones examined. The other viral antigens were not detected. The surface antigen was excreted into the cell culture medium as particles having the same characteristics as those found in human serum. It is estimated that 2-4 X 10' particles were produced per mouse cell per 24 hr in two clones. This value corresponds to approximately24 X 106 surface antigen polypeptides per cell per 24 hr.Hepatitis B virus (HBV) has a limited host range and, in nature, seems to infect only man and perhaps a few additional primates (1). So far, the virus has not been propagated in cell culture, and some viral surface antigen-producing cell lines from human hepatocellular carcinomas constitute the only cell culture systems that synthesize a HBV marker (2,3). For these reasons, study of the virus multiplication at the molecular level has been greatly hampered. Recently, information on the genetic organization of the virus was obtained from the nucleotide sequence of the genome. Genes coding for surface and capsid antigens and perhaps for DNA polymerase are located on the long strand of the genome (4-7). However, nothing is known concerning virus gene expression and its regulation.One approach to the study of HBV gene expression is to examine the functional capacity of the cloned viral DNA after its introduction into a mammalian cell culture. A gene that does not code for a selectable marker can be transferred into a cell by cotransformation of mutant mouse L cells deficient in thymidine kinase (TK) with this gene and the herpes simplex virus (HSV) tk gene (8). It appears that most of the selected.TK+ colonies contain the nonselectable marker. In the present study, mouse LTK-cells were cotransformed with HBV cloned DNA and the HSV cloned tk gene. All the selected TK+ clones synthesized hepatitis B surface antigen (HBsAg) particles which were excreted into the culture medium without apparent damage to the cells.MATERIALS AND METHODS Construction of the Recombinant and Bacterial Transformation. Plasmid pBR322 (200 ng) was digested with EcoRI endonuclease and treated with 2.6 units of alkaline phosphatase (9) in 100 mM Tris'HCI (pH 8.0) at 600C for 60 min. After two phenol extractions and three ether extractions, DNA was precipitated with ethanol. The pellet was dissolved in water and 100 ng of EcoRI-digested HBV DNA was added. The ligation was performed as described (10). Escherichia coli DP50. was grown in L broth medium containing 100.ug of diamninopimelic acid (Sigma) and 20 ug of thymidine (Sigma) per ml. Bacteria were transformed as described (10) with the ligation mixture and were select...

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Localization on the viral genome and nucleotide sequence of the gene coding for the two major polypeptides of the Hepatitis B surface antigen (HBs Ag)

Cited by 74 publications

References 29 publications

Isolation, characterization, and comparison of recombinant DNAs derived from genomes of human hepatitis B virus and woodchuck hepatitis virus.

Isolation, characterization, and comparison of recombinant DNAs derived from genomes of human hepatitis B virus and woodchuck hepatitis virus.

Detection of Antiviral Antibodies with Predetermined Specificity Using Synthetic Peptide-beta-Lactamase Conjugates: Application to Antibodies Specific for the preS Region of the Hepatitis B Virus Envelope Proteins

Excretion of hepatitis B surface antigen particles from mouse cells transformed with cloned viral DNA.

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