Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1985 Jan;53(1):316–318. doi: 10.1128/jvi.53.1.316-318.1985

Mapping of a vaccinia host range sequence by insertion into the viral thymidine kinase gene.

S Gillard, D Spehner, R Drillien
PMCID: PMC255043  PMID: 2981355

Abstract

A vaccinia virus mutant deleted of ca. 18 kilobase pairs at the left-hand end of the genome is unable to multiply on many human cell lines. To determine whether all or some of the deleted sequences were responsible for the host range property, the corresponding region from wild-type DNA was cloned in three pieces into a vaccinia transplacement vector containing the thymidine kinase gene on the HindIII J fragment. The next step was to transfer these pieces to the genome of the host range deletion mutant by in vivo homologous recombination around the thymidine kinase locus. Transfer of one 5.2-kilobase-pair EcoRI fragment was found to restore a wild-type phenotype on the host range mutant, thus demonstrating that only a small portion of the 18-kilobase-pair deletion contains the host range function(s). This result also illustrates that the method initially devised for inserting foreign genes into vaccinia virus DNA is useful for studies of the vaccinia genome.

Full text

PDF
316

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Drillien R., Koehren F., Kirn A. Host range deletion mutant of vaccinia virus defective in human cells. Virology. 1981 Jun;111(2):488–499. doi: 10.1016/0042-6822(81)90351-2. [DOI] [PubMed] [Google Scholar]
  2. Drillien R., Spehner D. Physical mapping of vaccinia virus temperature-sensitive mutations. Virology. 1983 Dec;131(2):385–393. doi: 10.1016/0042-6822(83)90506-8. [DOI] [PubMed] [Google Scholar]
  3. Fenner F., Sambrook J. F. Conditional lethal mutants of rabbitpox virus. II. Mutants (p) that fail to multiply in PK-2a cells. Virology. 1966 Apr;28(4):600–609. doi: 10.1016/0042-6822(66)90245-5. [DOI] [PubMed] [Google Scholar]
  4. GEMMELL A., FENNER F. Genetic studies with mammalian poxviruses. III. White (u) mutants of rabbitpox virus. Virology. 1960 May;11:219–235. doi: 10.1016/0042-6822(60)90063-5. [DOI] [PubMed] [Google Scholar]
  5. Isle H. B., Venkatesan S., Moss B. Cell-free translation of early and late mRNAs selected by hybridization to cloned DNA fragments derived from the left 14 million to 72 million daltons of the vaccinia virus genome. Virology. 1981 Jul 15;112(1):306–317. doi: 10.1016/0042-6822(81)90636-x. [DOI] [PubMed] [Google Scholar]
  6. Lake J. R., Cooper P. D. Deletions of the terminal sequences in the genomes of the white pock (u) and host-restricted (p) mutants of rabbitpox virus. J Gen Virol. 1980 May;48(1):135–147. doi: 10.1099/0022-1317-48-1-135. [DOI] [PubMed] [Google Scholar]
  7. Mackett M., Smith G. L., Moss B. Vaccinia virus: a selectable eukaryotic cloning and expression vector. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7415–7419. doi: 10.1073/pnas.79.23.7415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Moss B., Winters E., Cooper J. A. Deletion of a 9,000-base-pair segment of the vaccinia virus genome that encodes nonessential polypeptides. J Virol. 1981 Nov;40(2):387–395. doi: 10.1128/jvi.40.2.387-395.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Moyer R. W., Rothe C. T. The white pock mutants of rabbit poxvirus. I. Spontaneous host range mutants contain deletions. Virology. 1980 Apr 15;102(1):119–132. doi: 10.1016/0042-6822(80)90075-6. [DOI] [PubMed] [Google Scholar]
  10. Panicali D., Davis S. W., Mercer S. R., Paoletti E. Two major DNA variants present in serially propagated stocks of the WR strain of vaccinia virus. J Virol. 1981 Mar;37(3):1000–1010. doi: 10.1128/jvi.37.3.1000-1010.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Sambrook J. F., Padgett B. L., Tomkins J. K. Conditional lethal mutants of rabbitpox virus. I. Isolation of host cell-dependent and temperature-dependent mutants. Virology. 1966 Apr;28(4):592–599. doi: 10.1016/0042-6822(66)90244-3. [DOI] [PubMed] [Google Scholar]
  12. TAGAYA I., KITAMURA T., SANO Y. A new mutant of dermovaccinia virus. Nature. 1961 Oct 28;192:381–382. doi: 10.1038/192381a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES