Generation of varicella-zoster virus (VZV) and viral mutants from cosmid DNAs: VZV thymidylate synthetase is not essential for replication in vitro.
- 1 August 1993
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 90 (15), 7376-7380
- https://doi.org/10.1073/pnas.90.15.7376
Abstract
Four overlapping cosmid clones were constructed that contain the complete genome of the attenuated Oka strain of VZV. Transfection of human melanoma cells with the four cosmids resulted in production of infectious VZV. A double-stranded oligonucleotide, encoding a stop codon in all three open reading frames, was inserted into one of the cosmids at the 5' end of the viral thymidylate synthetase gene. Transfection of melanoma cells with the mutant cosmid, along with the other three cosmids, resulted in VZV that does not express the viral thymidylate synthetase protein. The mutant virus grew at a rate similar to that of the parental Oka strain virus. Production of recombinant VZV using cosmid DNAs will be useful for studying the function of viral genes in VZV replication and establishment of latency. Furthermore, manipulation of the Oka strain of VZV might allow one to produce a vaccine virus that does not establish latency in the central nervous system or a virus that encodes foreign antigens for use as a polyvalent live virus vaccine.Keywords
This publication has 14 references indexed in Scilit:
- Regulation of varicella-zoster virus gene expression in human T lymphocytesJournal of Virology, 1992
- Development of immunogenic recombinant Oka varicella vaccine expressing hepatitis B virus surface antigenJournal of General Virology, 1991
- Varicella-zoster virusJournal of General Virology, 1991
- Investigation of varicella-zoster virus infection of lymphocytes by in situ hybridizationJournal of Virology, 1989
- 2-Acetylpyridine 5-[(dimethylamino)thiocarbonyl]-thiocarbonohydrazone (A1110U), a potent inactivator of ribonucleotide reductases of herpes simplex and varicella-zoster viruses and a potentiator of acyclovir.Proceedings of the National Academy of Sciences, 1989
- Randomly picked cosmid clones overlap thepyrB andoriC gap in the physical map of theE.colichromosomeNucleic Acids Research, 1988
- Varicella-zoster virus as a live vector for the expression of foreign genes.Proceedings of the National Academy of Sciences, 1987
- Varicella-Zoster Virus Specifies a Thymidylate SynthetaseJournal of General Virology, 1987
- The Complete DNA Sequence of Varicella-Zoster VirusJournal of General Virology, 1986
- The A+T-rich genome of Herpesvirus saimiri contains a highly conserved gene for thymidylate synthase.Proceedings of the National Academy of Sciences of the United States of America, 1986