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. 1983 Nov;48(2):419–428. doi: 10.1128/jvi.48.2.419-428.1983

Marker rescue of temperature-sensitive mutations of vaccinia virus WR: correlation of genetic and physical maps.

M J Ensinger, M Rovinsky
PMCID: PMC255367  PMID: 6312100

Abstract

The physical map locations of 62 temperature-sensitive mutations of vaccinia virus WR have been determined by marker rescue experiments, using cloned HindIII fragments of wild-type DNA. Since vaccinia virus DNA is not infectious, marker rescue was performed by infecting monolayers of cells at the nonpermissive temperature with a low multiplicity of the mutant to be rescued and transfecting with calcium phosphate-precipitated recombinant DNA. Wild-type recombinants were measured by using either a direct plaque assay technique or a two-step procedure in which the final yield of virus from the transfected cells was assayed at the permissive and nonpermissive temperatures. Mutants that had been previously assigned to the same complementation-recombination group were rescued by the same HindIII fragment, with the exception of three mutants in one group that were rescued by either one of two adjacent fragments. A comparison between the genetic linkage map of the temperature-sensitive mutations in 30 mutants with their physical locations demonstrated that not only was the order of the genetic map correct but also recombination frequencies generally reflected actual physical distances.

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Selected References

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

  1. Bajszár G., Wittek R., Weir J. P., Moss B. Vaccinia virus thymidine kinase and neighboring genes: mRNAs and polypeptides of wild-type virus and putative nonsense mutants. J Virol. 1983 Jan;45(1):62–72. doi: 10.1128/jvi.45.1.62-72.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brockman W. W., Nathans D. The isolation of simian virus 40 variants with specifically altered genomes. Proc Natl Acad Sci U S A. 1974 Mar;71(3):942–946. doi: 10.1073/pnas.71.3.942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cabrera C. V., Esteban M., McCarron R., McAllister W. T., Holowczak J. A. Vaccinia virus transcription: hybridization of mRNA to restriction fragments of vaccinia DNA. Virology. 1978 May 1;86(1):102–114. doi: 10.1016/0042-6822(78)90011-9. [DOI] [PubMed] [Google Scholar]
  5. Chernos V. I., Belanov E. F., Vasilieva N. N. Temperature-sensitive mutants of vaccinia virus. I. Isolation and preliminary characterization. Acta Virol. 1978 Mar;22(2):81–90. [PubMed] [Google Scholar]
  6. Chipchase M., Schwendimann F., Wyler R. A map of the late proteins of vaccinia virus. Virology. 1980 Aug;105(1):261–264. doi: 10.1016/0042-6822(80)90176-2. [DOI] [PubMed] [Google Scholar]
  7. Condit R. C., Motyczka A. Isolation and preliminary characterization of temperature-sensitive mutants of vaccinia virus. Virology. 1981 Aug;113(1):224–241. doi: 10.1016/0042-6822(81)90150-1. [DOI] [PubMed] [Google Scholar]
  8. Cooper J. A., Moss B. Translation of specific vaccinia virus RNAs purified as RNA-DNA hybrids on potassium iodide gradients. Nucleic Acids Res. 1979 Aug 10;6(11):3599–3612. doi: 10.1093/nar/6.11.3599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cooper J. A., Wittek R., Moss B. Extension of the transcriptional and translational map of the left end of the vaccinia virus genome to 21 kilobase pairs. J Virol. 1981 Sep;39(3):733–745. doi: 10.1128/jvi.39.3.733-745.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cooper J. A., Wittek R., Moss B. Hybridization selection and cell-free translation of mRNA's encoded within the inverted terminal repetition of the vaccinia virus genome. J Virol. 1981 Jan;37(1):284–294. doi: 10.1128/jvi.37.1.284-294.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dales S., Milovanovitch V., Pogo B. G., Weintraub S. B., Huima T., Wilton S., McFadden G. Biogenesis of vaccinia: isolation of conditional lethal mutants and electron microscopic characterization of their phenotypically expressed defects. Virology. 1978 Feb;84(2):403–428. doi: 10.1016/0042-6822(78)90258-1. [DOI] [PubMed] [Google Scholar]
  12. DeFilippes F. M. Restriction enzyme mapping of vaccinia virus DNA. J Virol. 1982 Jul;43(1):136–149. doi: 10.1128/jvi.43.1.136-149.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Drillien R., Spehner D., Kirn A. Complementation and genetic linkage between vaccinia virus temperature-sensitive mutants. Virology. 1982 Jun;119(2):372–381. doi: 10.1016/0042-6822(82)90096-4. [DOI] [PubMed] [Google Scholar]
  14. Drillien R., Tripier F., Koehren F., Kirn A. A temperature-sensitive mutant of vaccinia virus defective in an early stage of morphogenesis. Virology. 1977 Jun 15;79(2):369–380. doi: 10.1016/0042-6822(77)90364-6. [DOI] [PubMed] [Google Scholar]
  15. Ensinger M. J. Isolation and genetic characterization of temperature-sensitive mutants of vaccinia virus WR. J Virol. 1982 Sep;43(3):778–790. doi: 10.1128/jvi.43.3.778-790.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Lake J. R., Silver M., Dales S. Biogenesis of vaccinia: complementation and recombination analysis of one group of conditional-lethal mutants defective in envelope self-assembly. Virology. 1979 Jul 15;96(1):9–20. doi: 10.1016/0042-6822(79)90167-3. [DOI] [PubMed] [Google Scholar]
  19. Mackett M., Archard L. C. Conservation and variation in Orthopoxvirus genome structure. J Gen Virol. 1979 Dec;45(3):683–701. doi: 10.1099/0022-1317-45-3-683. [DOI] [PubMed] [Google Scholar]
  20. Nakano E., Panicali D., Paoletti E. Molecular genetics of vaccinia virus: demonstration of marker rescue. Proc Natl Acad Sci U S A. 1982 Mar;79(5):1593–1596. doi: 10.1073/pnas.79.5.1593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Padgett B. L., Tomkins J. K. Conditional lethal mutants of rabbitpox virus. 3. Temperature-sensitive (ts) mutants; physiological properties, complementation and recombination. Virology. 1968 Oct;36(2):161–167. doi: 10.1016/0042-6822(68)90131-1. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Weir J. P., Bajszár G., Moss B. Mapping of the vaccinia virus thymidine kinase gene by marker rescue and by cell-free translation of selected mRNA. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1210–1214. doi: 10.1073/pnas.79.4.1210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wittek R., Barbosa E., Cooper J. A., Garon C. F., Chan H., Moss B. Inverted terminal repetition in vaccinia virus DNA encodes early mRNAs. Nature. 1980 May 1;285(5759):21–25. doi: 10.1038/285021a0. [DOI] [PubMed] [Google Scholar]
  25. Wittek R., Cooper J. A., Barbosa E., Moss B. Expression of the vaccinia virus genome: analysis and mapping of mRNAs encoded within the inverted terminal repetition. Cell. 1980 Sep;21(2):487–493. doi: 10.1016/0092-8674(80)90485-7. [DOI] [PubMed] [Google Scholar]
  26. Wittek R., Cooper J. A., Moss B. Transcriptional and translational mapping of a 6.6-kilobase-pair DNA fragment containing the junction of the terminal repetition and unique sequence at the left end of the vaccinia virus genome. J Virol. 1981 Sep;39(3):722–732. doi: 10.1128/jvi.39.3.722-732.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

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