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Growth and Virulence Alterations of Equine Herpesvirus 1 by Insertion of a Green Fluorescent Protein Gene in the Intergenic Region between ORFs 62 and 63
Author(s) -
Ibrahim El Sayed Moustafa,
Pagmajav Ochir,
Yamaguchi Tsuyoshi,
Matsumura Tomio,
Fukushi Hideto
Publication year - 2004
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.2004.tb03615.x
Subject(s) - orfs , biology , green fluorescent protein , intergenic region , mutant , virulence , gene , virology , genetics , wild type , microbiology and biotechnology , virus , open reading frame , genome , peptide sequence
Nucleotide sequences of the intergenic region between ORF 62 and ORF 63 of equine herpesvirus 1 (EHV‐1) isolates were analyzed. The sequences of this region consisted of variable and conserved domains among EHV‐1 isolates. An EHV‐1 mutant, Ab4‐GFP, was constructed by inserting a green fluorescent protein (GFP) expression cassette flanked by lox P at both ends into the intergenic region between ORF 62 and ORF 63. Another mutant, Ab4‐loxP, which contains one lox P site, was constructed by excision of the GFP cassette from the Ab4‐GFP virus genome by cre enzyme. The recombinant Ab4‐GFP formed smaller plaques than the wild type in MDBK cells. Virus production also decreased for Ab4‐GFP in multi‐step growth analyses. Virulence of Ab4‐GFP in both mice and hamsters was weaker than that of the wild type. Ab4‐loxP exhibited properties similar to those of the wild type. These results suggest that the intergenic region between ORF 62 and ORF 63 plays various roles in the virus growth.