Cloning of the varicella-zoster virus genome as an infectious bacterial artificial chromosome in Escherichia coli.
The complete genome of the varicella-zoster virus (VZV) Oka strain has been cloned as a bacterial artificial chromosome (BAC). Following electroporation into Escherichia coli (E. coli) strain DH10B, the VZV BAC was stably propagated over multiple generations of its host. Human embryonic lung (HEL) cells transfected with VZV BAC DNA recovered from DH10B showed cytopathic effect (CPE), and virus spread to neighbouring cells was observed. BAC vector sequences are flanked by loxP sites and, coinfection of the reconstituted virus, with a recombinant adenovirus expressing Cre recombinase removed the bacterial sequences. The resulting recombinant rV02 grew as well as the parental virus in HEL cells. The recombinant VZV will promote VZV research and increase use of the viral genome as an investigative tool.
Nagaike
Kazuhiro
K
Department of Microbiology, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita, Osaka 565-0871, Japan.
Mori
Yasuko
Y
Gomi
Yasuyuki
Y
Yoshii
Hironori
H
Takahashi
Michiaki
M
Wagner
Markus
M
Koszinowski
Ulrich
U
Yamanishi
Koichi
K
D000256:Adenoviridae / Q000235:genetics; D002478:Cells, Cultured; D022202:Chromosomes, Artificial, Bacterial; D003001:Cloning, Molecular; D003588:Cytopathogenic Effect, Viral; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D005822:Genetic Vectors / Q000235:genetics; D016679:Genome, Viral; D014645:Herpesvirus 3, Human / Q000235:genetics* / Q000502:physiology; D006801:Humans; D045522:Recombinases / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014779:Virus Replication
D004279:DNA, Viral; D045522:Recombinases