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Identification, cloning, and expression of the major capsid protein gene of human herpesvirus 6
E Littler1, G Lawrence, M Y Liu
1Department of Molecular Biology, Christie Hospital & Holt Radium Institute, Manchester, United Kingdom.
Abstract:
DNA sequence analysis of part of the human herpesvirus 6 (HHV-6) genome led to the identification of an open reading frame with amino acid sequence homology to the major capsid proteins (MCP) of other HHVs. DIAGON analysis showed that the closest homology was with human cytomegalovirus. Plasmids were constructed which were shown to express the HHV-6 MCP as either the entire open reading frame or as portions of it, and the recombinant-produced proteins were used to raise antisera. The antisera were shown by immunofluorescence to react with HHV-6-infected lymphoblastoid cells and in Western blots with a 135-kilodalton protein specific to HHV-6-infected cells. The recombinant protein expressed from the entire HHV-6 MCP gene was detected only weakly in Western blot assays with normal HHV-6-positive human sera as a probe.
Insights
Researchers identified a gene in human herpesvirus 6 (HHV-6) encoding a protein similar to major capsid proteins (MCP) of other herpesviruses. This discovery aids in understanding HHV-6 structure and potential therapeutic targets.
Area of Science:
- Virology
- Molecular Biology
- Genomics
Background:
- Human herpesvirus 6 (HHV-6) is a significant human pathogen.
- Understanding the HHV-6 genome, particularly its structural proteins, is crucial for viral research.
Purpose of the Study:
- To identify and characterize the gene encoding the major capsid protein (MCP) of HHV-6.
- To develop tools for detecting HHV-6 infection and studying its viral components.
Main Methods:
- DNA sequence analysis of the HHV-6 genome.
- Homology searches using DIAGON analysis.
- Plasmid construction for recombinant protein expression.
- Antibody production (antisera) against recombinant HHV-6 MCP.
- Immunofluorescence and Western blot assays for protein detection.
Main Results:
- An open reading frame homologous to HHV MCPs was identified in the HHV-6 genome.
- The closest homology was found with human cytomegalovirus MCP.
- Recombinant HHV-6 MCP was expressed and used to generate specific antisera.
- Antisera recognized a 135-kDa protein in HHV-6 infected cells via immunofluorescence and Western blot.
Conclusions:
- The identified HHV-6 gene encodes a protein functionally analogous to the major capsid protein of other human herpesviruses.
- This finding provides a new target for HHV-6 diagnostics and antiviral development.
- Further characterization of this protein is warranted for a comprehensive understanding of HHV-6 virion structure.