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March 1, 1993Journal of VirologyOpen Access

Cloning and characterization of human astrovirus immunoreactive epitopes

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Authors

SMS M MatsuiJKJ P KimHGHarry B. Greenberg

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Overview

Laboratory investigation identifies cross-reactive astrovirus epitopes using expression cloning, indicating that viral antigens are encoded across genomic and subgenomic RNAs.

Key Points

  • To identify and clone conserved, immunoreactive protein-coding regions across human astrovirus serotypes.
  • Screened human astrovirus serotype 1 lambda gt11 cDNA libraries using hyperimmune rabbit antiserum generated against purified astrovirus particles.
  • Evaluated clone specificity using preimmunization and postimmunization rabbit sera.
  • Analyzed viral transcript origins by hybridizing radiolabeled cDNA clone probes to astrovirus genomic and subgenomic RNAs.
  • Identified four astrovirus-specific clones consisting of three overlapping clones (A43, A35, and A1) and one distinct clone (A14) that reacted specifically with immune serum.
  • Clones A43-A35-A1 hybridized exclusively to the 7.2-kb full-length genomic RNA.
  • Clone A14 hybridized to both the 7.2-kb genomic RNA and the 2.8-kb subgenomic RNA, indicating epitope coding regions span both RNA species.

Cite This Study

Matsui et al. (1993) studied this question.

synapsesocial.com/papers/6a6fb062febe604dd7088d29https://doi.org/10.1128/jvi.67.3.1712-1715.1993
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