scholarly journals Functional Profiling of Varicella-Zoster Virus Genome by Use of a Luciferase Bacterial Artificial Chromosome System

10.5772/31838 ◽  
2011 ◽  
Author(s):  
Lucy Zhu ◽  
Hua Zhu
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Varicella Zoster Virus ◽  
Artificial Chromosome ◽  
Virus Genome ◽  
Varicella Zoster ◽  
Functional Profiling

scholarly journals Genetic Analysis of Varicella-Zoster Virus ORF0 to ORF4 by Use of a Novel Luciferase Bacterial Artificial Chromosome System

2007 ◽  
Vol 81 (17) ◽  
pp. 9024-9033 ◽  
Author(s):  
Zhen Zhang ◽  
Jenny Rowe ◽  
Weijia Wang ◽  
Marvin Sommer ◽  
Ann Arvin ◽  
...  
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Varicella Zoster Virus ◽  
Artificial Chromosome ◽  
Wild Type Virus ◽  
Growth Curve Analysis ◽  
Type Virus ◽  
Wild Type ◽  
Varicella Zoster

ABSTRACT To efficiently generate varicella-zoster virus (VZV) mutants, we inserted a bacterial artificial chromosome (BAC) vector in the pOka genome. We showed that the recombinant VZV (VZVBAC) strain was produced efficiently from the BAC DNA and behaved indistinguishably from wild-type virus. Moreover, VZV's cell-associated nature makes characterizing VZV mutant growth kinetics difficult, especially when attempts are made to monitor viral replication in vivo. To overcome this problem, we then created a VZV strain carrying the luciferase gene (VZVLuc). This virus grew like the wild-type virus, and the resulting luciferase activity could be quantified both in vitro and in vivo. Using PCR-based mutagenesis, open reading frames (ORF) 0 to 4 were individually deleted from VZVLuc genomes. The deletion mutant viruses appeared after transfection into MeWo cells, except for ORF4, which was essential. Growth curve analysis using MeWo cells and SCID-hu mice indicated that ORF1, ORF2, and ORF3 were dispensable for VZV replication both in vitro and in vivo. Interestingly, the ORF0 deletion virus showed severely retarded growth both in vitro and in vivo. The growth defects of the ORF0 and ORF4 mutants could be fully rescued by introducing wild-type copies of these genes back into their native genome loci. This work has validated and justified the use of the novel luciferase VZV BAC system to efficiently generate recombinant VZV variants and ease subsequent viral growth kinetic analysis both in vitro and in vivo.

scholarly journals A Self-Excisable Infectious Bacterial Artificial Chromosome Clone of Varicella-Zoster Virus Allows Analysis of the Essential Tegument Protein Encoded by ORF9

2007 ◽  
Vol 81 (23) ◽  
pp. 13200-13208 ◽  
Author(s):  
B. Karsten Tischer ◽  
Benedikt B. Kaufer ◽  
Marvin Sommer ◽  
Felix Wussow ◽  
Ann M. Arvin ◽  
...  
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Varicella Zoster Virus ◽  
Recombinant Virus ◽  
Artificial Chromosome ◽  
Full Length ◽  
Start Codon ◽  
Tegument Protein ◽  
Growth Defect ◽  
Bac Clone ◽  
Varicella Zoster

ABSTRACT In order to facilitate the generation of mutant viruses of varicella-zoster virus (VZV), the agent causing varicella (chicken pox) and herpes zoster (shingles), we generated a full-length infectious bacterial artificial chromosome (BAC) clone of the P-Oka strain. First, mini-F sequences were inserted into a preexisting VZV cosmid, and the SuperCos replicon was removed. Subsequently, mini-F-containing recombinant virus was generated from overlapping cosmid clones, and full-length VZV DNA recovered from the recombinant virus was established in Escherichia coli as an infectious BAC. An inverted duplication of VZV genomic sequences within the mini-F replicon resulted in markerless excision of vector sequences upon virus reconstitution in eukaryotic cells. Using the novel tool, the role in VZV replication of the major tegument protein encoded by ORF9 was investigated. A markerless point mutation introduced in the start codon by two-step en passant Red mutagenesis abrogated ORF9 expression and resulted in a dramatic growth defect that was not observed in a revertant virus. The essential nature of ORF9 for VZV replication was ultimately confirmed by restoration of the growth of the ORF9-deficient mutant virus using trans-complementation via baculovirus-mediated gene transfer.

scholarly journals Inversion and circularization of the varicella-zoster virus genome.

1985 ◽  
Vol 56 (1) ◽  
pp. 194-200 ◽  
Author(s):  
P R Kinchington ◽  
W C Reinhold ◽  
T A Casey ◽  
S E Straus ◽  
J Hay ◽  
...  
Keyword(s):  
Varicella Zoster Virus ◽  
Virus Genome ◽  
Varicella Zoster

scholarly journals Transcription mapping of the varicella-zoster virus genome.

1985 ◽  
Vol 56 (2) ◽  
pp. 600-606 ◽  
Author(s):  
J M Ostrove ◽  
W Reinhold ◽  
C M Fan ◽  
S Zorn ◽  
J Hay ◽  
...  
Keyword(s):  
Varicella Zoster Virus ◽  
Virus Genome ◽  
Varicella Zoster

scholarly journals XbaI, PstI, and BglII restriction enzyme maps of the two orientations of the varicella-zoster virus genome.

1981 ◽  
Vol 39 (2) ◽  
pp. 390-400 ◽  
Author(s):  
A M Dumas ◽  
J L Geelen ◽  
M W Weststrate ◽  
P Wertheim ◽  
J van der Noordaa
Keyword(s):  
Varicella Zoster Virus ◽  
Restriction Enzyme ◽  
Virus Genome ◽  
Varicella Zoster ◽  
Restriction Enzyme Maps

scholarly journals Characterization of Varicella-Zoster Virus Gene 21 and 29 Proteins in Infected Cells

2002 ◽  
Vol 76 (14) ◽  
pp. 7228-7238 ◽  
Author(s):  
Randall J. Cohrs ◽  
Jeanne Wischer ◽  
Carrie Essman ◽  
Donald H. Gilden
Keyword(s):  
Varicella Zoster Virus ◽  
Virus Genome ◽  
Open Reading Frames ◽  
Productive Infection ◽  
Gene Promoters ◽  
Varicella Zoster ◽  
Latently Infected ◽  
Infected Cells ◽  
Simplex Virus ◽  
Hsv 1

ABSTRACT Varicella-zoster virus (VZV) transcription is limited in latently infected human ganglia. Note that much of the transcriptional capacity of the virus genome has not been analyzed in detail; to date, only VZV genes mapping to open reading frames (ORFs) 4, 21, 29, 62, and 63 have been detected. ORF 62 encodes the major immediate-early virus transcription transactivator IE62, ORF 29 encodes the major virus DNA binding protein, and ORF 21 encodes a protein associated with the developing virus nucleocapsid. We analyzed the cellular location of proteins encoded by ORF 21 (21p) and ORF 29 (29p), their phosphorylation state during productive infection, and their ability form a protein-protein complex. The locations of both 21p and 29p within infected cells mimic those of their herpes simplex virus type 1 (HSV-1) homologues (UL37 and ICP8); however, unlike these homologues, 21p is not phosphorylated and neither 21p nor 29p exhibits a protein-protein interaction. Transient transfection assays to determine the effect of 21p and 29p on transcription from VZV gene 20, 21, 28, and 29 promoters revealed no significant activation of transcription by 21p or 29p from any of the VZV gene promoters tested, and 21p did not significantly modulate the ability of IE62 to activate gene transcription. A modest increase in IE62-induced activation of gene 28 and 29 promoters was seen in the presence of 29p; however, IE62-induced activation of gene 28 and 29 promoters was reduced in the presence of 21p. A Saccharomyces cerevisiae two-hybrid analysis of 21p indicated that the protein can activate transcription when tethered within a responsive promoter. Together, the data reveal that while VZV gene 21 and HSV-1 UL37 share homology at the nucleic acid level, these proteins differ functionally.

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