Identification and analysis of an alcelaphine herpesvirus 1 (AHV-1) cDNA clone expressing a fusion protein recognized by AHV-1-neutralizing antisera.
Rabbit antiserum to psoralen-inactivated alcelaphine herpesvirus 1 (AHV-1) virions was shown to react specifically with AHV-1-infected cells by indirect immunofluorescence. Western blot analysis using this antiserum identified a 15-kD virion protein that was also detected in infected-cell proteins between 12 and 144 h p.i., and a 37-kD protein present in infected cells between 24 and 120 h p.i. A cDNA library was constructed using mRNA obtained from AHV-1-infected fetal mouflon sheep kidney (FMSK) cells at 48 h p.i., when infected-cell proteins detected by antiserum were in abundance. Screening of the library with the rabbit anti-AHV-1 serum identified several positive clones. Southern blot analysis showed that one clone, designated 8'a, hybridized to a 4.4 kb HindIII fragment of AHV-1 DNA. This AHV-1 cDNA clone expressed a fusion protein that was recognized by serum from a naturally and asymptomatically infected white-bearded wildebeest (Connochaetes taurinus albojubatus). The insert was sequenced and found to contain 833 bp. A search of the GenBank database for related sequences revealed greater than 40% homology to several other gammaherpesviruses: herpesvirus saimiri, cottontail herpesvirus, and Epstein-Barr virus.
Lahijani
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Center for Reproduction of Endangered Species, Zoological Society of San Diego, California, USA.
Sutton
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SM
Klieforth
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Heuschele
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WP
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D018076:DNA, Complementary / Q000302:isolation & purification*; D005455:Fluorescent Antibody Technique; D006564:Herpesviridae / Q000235:genetics* / Q000276:immunology; D007106:Immune Sera / Q000276:immunology*; D008969:Molecular Sequence Data; D011817:Rabbits; D012756:Sheep; D014760:Viral Fusion Proteins / Q000235:genetics* / Q000276:immunology
D000956:Antigens, Viral; D018076:DNA, Complementary; D007106:Immune Sera; D014760:Viral Fusion Proteins