Construction of a viable BHV1 mutant lacking most of the short unique region.
The 8.2 kb HindIII K-fragment of bovine herpesvirus 1 (BHV1) lies entirely within the short unique region of the genome and contains all or parts of 7 coding regions. We have probed this fragment for the presence of nonessential genes by a simple, rapid method of insertional mutagenesis. Our analysis indicates that all the genes present in the K-fragment, except the glycoprotein D gene, are nonessential for replication of BHV1 in tissue culture. After inserting a copy of the glycoprotein D gene into the thymidine kinase locus of BHV1 it was possible to delete the entire HindIII K-fragment and the contiguous 0.36 kb O-fragment in one step. The deletion mutant, which contains 7 kb less BHV1 DNA than wt virus and lacks ORF1, US2, the genes for protein kinase, glycoprotein G, glycoprotein I, and most of glycoprotein E was still replication-competent.
Department of Pathology and Laboratory Medicine, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
J J
JJ
Furth
J C
JC
Whitbeck
W C
WC
Lawrence
L J
LJ
Bello
D000818:Animals; D002417:Cattle; D002460:Cell Line; D002874:Chromosome Mapping; D015247:Deoxyribonuclease HindIII / Q000378:metabolism; D007242:Herpesvirus 1, Bovine / Q000235:genetics* / Q000254:growth & development / Q000502:physiology*; D016254:Mutagenesis, Insertional; D011995:Recombination, Genetic; D014764:Viral Proteins / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D014764:Viral Proteins; C046998:bovine herpesvirus type-1 glycoproteins; D015247:Deoxyribonuclease HindIII; D001616:beta-Galactosidase