A highly efficient protocol of generating and analyzing VZV ORF deletion mutants based on a newly developed luciferase VZV BAC system.
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.
Zhang
Zhen
Z
Department of Microbiology and Molecular Genetics, UMDNJ-New Jersey Medical School, 225 Warren Street, Newark, NJ 07101-1709, USA.
Huang
Ying
Y
Zhu
Hua
H
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*
D049409:Luciferases, Firefly