Identification and localization of pre-s-encoded polypeptides from woodchuck and ground squirrel hepatitis viruses
| Autor: | E Schaeffer, R L Snyder, J J Sninsky |
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| Rok vydání: | 1986 |
| Předmět: |
Genes
Viral Hepatitis B virus DNA polymerase viruses Genetic Vectors Immunology DNA-Directed DNA Polymerase Hepatitis Animal Microbiology Virus law.invention Rodent Diseases Chloramphenicol acetyltransferase Viral Proteins law Virology Hepatitis Viruses Animals chemistry.chemical_classification Antiserum biology Woodchuck hepatitis virus Virion Sciuridae biology.organism_classification Fusion protein Molecular biology Recombinant Proteins Amino acid chemistry Marmota Insect Science Recombinant DNA Plasmids Research Article |
| Zdroj: | Journal of Virology. 57:173-182 |
| ISSN: | 1098-5514 0022-538X |
| Popis: | A segment from the pre-s region of the woodchuck hepatitis virus (WHV) was inserted into an open reading frame vector allowing for the expression in Escherichia coli of viral determinants as part of a fusion protein. The bacterially synthesized fusion molecule contained eight amino acids from beta-galactosidase (beta-gal) at the N terminus, followed by 89 pre-s-encoded amino acids and 219 amino acids of chloramphenicol acetyltransferase (CAT) at the C terminus (beta-gal:pre-s:CAT). This tribrid protein was used to generate antiserum which had a significant titer to the viral portion of the fusion polypeptide. Anti-beta-gal:pre-s:CAT was used in Western blot analysis to identify viral proteins containing pre-s-encoded determinants. Antiserum to the tribrid molecule recognized four WHV polypeptides with molecular masses of 33, 36, 45, and 47 kilodaltons, each of which was also recognized by a monoclonal antibody to WHV surface antigen. Using the same anti-tribrid serum, we also identified analogous polypeptides from ground squirrel hepatitis virus. The antiserum was also used to immunoprecipitate virus particles containing endogenous DNA polymerase activity, indicating that pre-s determinants are found on the surface of mature virions. Based on previous computer studies and the location of pre-s-encoded molecules on the surface of virus particles, a role in hepadnavirus host cell entry is suggested for these polypeptides. |
| Databáze: | OpenAIRE |
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