Construction of a viable BHV1 mutant lacking mostof the short unique region |
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Authors: | J J Furth J C Whitbeck W C Lawrence L J Bello |
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Institution: | (1) Department of Pathology and Laboratory Medicine, School of Medicine,;(2) Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, U.S.A., US |
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Abstract: | Summary. 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 glycoprotien E was still
replication-competent.
Accepted July 30, 1997 Received June 17, 1997 |
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