title: In vitro characterization of felid herpesvirus 1 (FHV-1) mutants generated by recombineering in a recombinant BAC vector.
abstract: Felid herpesvirus 1 (FHV-1) mutants were constructed using two-step Red-mediated recombination techniques based on a virulent full-length FHV-1 BAC clone. The individual mutant viruses generated were deficient in glycoprotein C (gC), glycoprotein E (gE), US3 serine/threonine protein kinase (PK), or both gE and thymidine kinase (TK). The gC- mutant virus produced plaques that were similar in size to those resulting from infection with the C-27 parent strain. In contrast, the gE(-), PK(-), and gE(-)PK(-) deletion mutants produced plaques that were significantly smaller. Multistep in vitro growth kinetics of the gE(-), PK(-), and gE(-)PK(-) viruses were slightly delayed compared to those of the C-27 parent strain. Peak progeny titers of these three mutants were approximately 10-fold lower than those generated with the C-27 strain. There was no delay in the growth kinetics of the gC- mutant, but the progeny virus titer obtained with this mutant was at least 3 logs lower compared to the parental strain titer. Based upon their in vitro characteristics, these mutants will be useful for the development of novel immunization strategies against this important feline pathogen.
authors.affiliation: Graduate Program in Comparative Medicine and Integrative Biology, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA; Diagnostic Center for Population and Animal Health, College of Veterinary Medicine, Michigan State University, 4125 Beaumont Road, Lansing, MI 48910, USA.
authors.forename: S-H Sheldon
authors.initials: SH
authors.lastname: Tai
authors.affiliation: Diagnostic Center for Population and Animal Health, College of Veterinary Medicine, Michigan State University, 4125 Beaumont Road, Lansing, MI 48910, USA.
authors.forename: Carine
authors.initials: C
authors.lastname: Holz
authors.affiliation: Department of Microbiology and Molecular Genetics, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Engstrom
authors.affiliation: United States Department of Agriculture, Agricultural Research Service, Avian Disease and Oncology Laboratory, 4279 E. Mount Hope Road, East Lansing, MI 48823, USA.
authors.forename: Hans H
authors.initials: HH
authors.lastname: Cheng
authors.affiliation: Graduate Program in Comparative Medicine and Integrative Biology, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA; Diagnostic Center for Population and Animal Health, College of Veterinary Medicine, Michigan State University, 4125 Beaumont Road, Lansing, MI 48910, USA; Department of Microbiology and Molecular Genetics, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA; Department of Pathobiology and Diagnostic Investigation, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA. Electronic address: Maes@dcpah.msu.edu.
authors.forename: Roger K
authors.initials: RK
authors.lastname: Maes
mesh_terms: D000818:Animals; D002415:Cats; D002460:Cell Line; D022202:Chromosomes, Artificial, Bacterial; D016296:Mutagenesis; D011995:Recombination, Genetic; D018140:Varicellovirus / Q000235:genetics* / Q000502:physiology*; D019562:Viral Load; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000235:genetics; D014779:Virus Replication
chemical_list: D014764:Viral Proteins
keywords: Bacterial artificial chromosome; Felid herpesvirus 1; Glycoprotein; Serine/threonine protein kinase; Thymidine kinase; Vaccine