title: A highly efficient protocol of generating and analyzing VZV ORF deletion mutants based on a newly developed luciferase VZV BAC system.
abstract: Varicella Zoster Virus (VZV) is the causative agent for both varicella (chicken pox) and herpes zoster (shingles). As a member of the human herpesvirus family, VZV contains a large DNA genome, encoding 70 unique open reading frames (ORFs). The functions of the majority of these ORFs remain unknown. Recently, the full-length VZV (P-Oka strain) genome was cloned as a VZV bacteria artificial chromosome (BAC) and additionally a firefly luciferase cassette was inserted to generate a novel luciferase VZV BAC. In this study, a highly efficient protocol has been developed exploiting the new luciferase VZV BAC system to rapidly isolate and characterize VZV ORF deletion mutants by growth curve analysis in cell culture.
authors.affiliation: Department of Microbiology and Molecular Genetics, UMDNJ-New Jersey Medical School, 225 Warren Street, Newark, NJ 07101-1709, USA.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhang
authors.forename: Ying
authors.initials: Y
authors.lastname: Huang
authors.forename: Hua
authors.initials: H
authors.lastname: Zhu
mesh_terms: D002460:Cell Line; D022202:Chromosomes, Artificial, Bacterial; D017353:Gene Deletion; D017930:Genes, Reporter; D005814:Genes, Viral; D014645:Herpesvirus 3, Human / Q000235:genetics* / Q000254:growth & development; D006801:Humans; D049409:Luciferases, Firefly / Q000096:biosynthesis / Q000235:genetics; D008967:Molecular Biology / Q000379:methods*; D010948:Viral Plaque Assay; D014773:Virology / Q000379:methods*
chemical_list: D049409:Luciferases, Firefly