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The ORF37 (UL24) is a neuropathogenicity determinant of equine herpesvirus 1 (EHV-1) in the mouse encephalitis model |
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| Authors: | Samy Kasem Souichi Yamada Tomio Matsumura Hanafy Madbouly Kenji Ohya |
| Affiliation: | a Department of Applied Veterinary Sciences, United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan b Laboratory of Veterinary Microbiology, Faculty of Applied Biological Sciences, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan c Molecular Biology Division, Epizootic Research Center, Equine Research Institute, Japan Racing Association, Shimotsuke, Tochigi 329-0412, Japan d The Avian Zoonosis Research Center, Faculty of Agriculture, Tottori University, 4-101 Koyama Minami, Tottori 680-8550, Japan |
| Abstract: | Equine herpesvirus 1 (EHV-1) bacterial artificial chromosome clone (Ab4p BAC) was established based on neuropathogenic strain Ab4p. ORF37 encoding UL24 was replaced with a selection cassette, rpsL-neo gene, to produce an ORF37 deletion mutant, Ab4p?ORF37. Transfection of RK-13 cells with Ab4p?ORF37 genome DNA produced infectious virus, indicating that ORF37 is not essential for EHV-1 replication in cell culture. Deletion of ORF37 had no effect on the transcript expression of neighboring genes, ORF36 and ORF38, and the growth activity in MDBK cells. Ab4pΔORF37 lost neuropathogenicity in CBA/N1 mice as indicated by the absence of any neurological disorders and death. The growth of Ab4pΔORF37 in cultivated neural cells was one order of magnitude lower than that of parental and revertant viruses. These results indicated that the ORF37 is a neuropathogenicity determinant of EHV-1 in the mouse encephalitis model. |
| Keywords: | BAC, bacterial artificial chromosome bp, base pair cDNA, complementary deoxyribonucleic acid DNA, deoxyribonucleic acid EHV-1, equine herpesvirus type 1 FBS, fetal bovine serum FEK, fetal equine kidney GFP, green fluorescent protein gp2, glycoprotein 2 ICP4, infected cell protein MDBK, Madin-Darby bovine kidney MEM, minimum essential medium MOI, multiplicity of infection NIH, National Institutes of Health nt, nucleotide ORF, open reading frame PCR, polymerase chain reaction pfu, plaque-forming unit RK-13, Rabbit kidney 13 RNA, ribonucleic acid RT-PCR, reverse transcription and polymerase chain reaction SPF, specific pathogen free SV40, simian virus 40 VZV, varicella zoster virus |
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