Alterations in the equine herpesvirus type-1 (EHV-1) strain RacH during attenuation.
The equine herpesvirus type-1 modified live-vaccine strain RacH (256th passage on porcine embryonic kidney cells) was investigated by restriction-enzyme analysis and compared to representative plaque isolates of the 12th passage (RacL11, RacL22) and 185th passage (RacM24, RacM36). The restriction patterns of all Rac plaque isolates differed compared with reference strain Ab4. The left UL terminus was shortened by 0.1 kbp and a missing BamHI site led to the fusion of the f and t fragments. In some Rac derivatives, losses of restriction sites without deletions were observed: 1. One BamHI site located in the ribosyl reductase gene was missing in RacH, RacM24, RacM36, and RacL22; and 2. An SalI site mapping to the gp14 (gB) gene was absent in RacM24, RacM36 and RacH. An identical deletion of 0.85 kbp in size was found in both copies of the inverted repeat (IR) regions of RacH. The deletion was present only in the terminal IR of the medium-passage derivative RacM36. By contrast, in the genomes of the apathogenic RacM24, as well as the pathogenic plaque isolates RacL11 and RacL22, no deletions in the IRs were detectable. Nucleotide-sequence and Northern-blot analyses revealed that the deletions led to the elimination of one or both copies of the gene 67 (IR6) open-reading frame in RacM36 and RacH and affected the gene 68 (EUS1) in RacH.
Hübert
P H
PH
Institute for Medical Microbiology, Infectious and Epidemic Diseases, Ludwig-Maximilian University of Munich, Germany.
Birkenmaier
S
S
Rziha
H J
HJ
Osterrieder
N
N
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002460:Cell Line; D006224:Cricetinae; D004279:DNA, Viral / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D005260:Female; D017353:Gene Deletion; D006566:Herpesviridae Infections / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary; D004861:Herpesvirus 1, Equid / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D006734:Horse Diseases / Q000276:immunology / Q000517:prevention & control; D006736:Horses; D007668:Kidney / Q000166:cytology / Q000196:embryology; D008647:Mesocricetus; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010641:Phenotype; D015183:Restriction Mapping; D013552:Swine; D014613:Vaccines, Attenuated / Q000235:genetics* / Q000276:immunology
D004279:DNA, Viral; D014613:Vaccines, Attenuated