Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1979 Nov;32(2):623–628. doi: 10.1128/jvi.32.2.623-628.1979

Physical mapping of a large-plaque mutation of adenovirus type 2.

G Chinnadurai, S Chinnadurai, J Brusca
PMCID: PMC353594  PMID: 501803

Abstract

We have developed a simple method based on cotransfection of overlapping DNA restriction fragments for construction of recombinants of adenovirus type 2 (Ad2) and Ad5. When Ad2 DNA digested with restriction endonuclease EcoRI was cotransfected with Ad5 DNA digested with SalI, recombination occurred between Ad2 EcoRI-A (map position 0 to 59) and Ad5 SalI-A (map position 45 to 100). Analysis of the recombinant DNAs by digestion with EcoRI or BamHI restriction endonucleases indicated that, as expected, recombination had occurred in overlapping sequences (map position 45 to 59) between the Ad2 EcoRI-A fragment and the Ad5 SalI-A fragment. By using this method, several recombinants were constructed between a large-plaque (lp) mutant of Ad2 and wild-type Ad5. Cleavage of the recombinant genomes with restriction endonucleases BamHI, EcoRI, and HindIII revealed that the lp mutation is located within the left 41% of Ad2 genome.

Full text

PDF
623

Images in this article

625Image
on p.625
626Image
on p.626
626Image
on p.626
627Image
on p.627

Selected References

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

  1. Arrand J. E. Mapping of adenovirus type 5 temperature-sensitive mutations by marker rescue in enhanced double DNA infections. J Gen Virol. 1978 Dec;41(3):573–586. doi: 10.1099/0022-1317-41-3-573. [DOI] [PubMed] [Google Scholar]
  2. Chinnadurai G., Chinnadurai S., Green M. Enhanced infectivity of adenovirus type 2 DNA and a DNA-protein complex. J Virol. 1978 Apr;26(1):195–199. doi: 10.1128/jvi.26.1.195-199.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chow L. T., Roberts J. M., Lewis J. B., Broker T. R. A map of cytoplasmic RNA transcripts from lytic adenovirus type 2, determined by electron microscopy of RNA:DNA hybrids. Cell. 1977 Aug;11(4):819–836. doi: 10.1016/0092-8674(77)90294-x. [DOI] [PubMed] [Google Scholar]
  4. Frost E., Williams J. Mapping temperature-sensitive and host-range mutations of adenovirus type 5 by marker rescue. Virology. 1978 Nov;91(1):39–50. doi: 10.1016/0042-6822(78)90353-7. [DOI] [PubMed] [Google Scholar]
  5. GREEN M., PINA M. BIOCHEMICAL STUDIES ON ADENOVIRUS MULTIPLICATION, VI. PROPERTIES OF HIGHLY PURIFIED TUMORIGENIC HUMAN ADENOVIRUSES AND THEIR DNA. Proc Natl Acad Sci U S A. 1964 Jun;51:1251–1259. doi: 10.1073/pnas.51.6.1251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. GREEN M., PINA M. Biochemical studies on adenovirus multiplication. IV. Isolation, purification, and chemical analysis of adenovirus. Virology. 1963 May;20:199–207. doi: 10.1016/0042-6822(63)90157-0. [DOI] [PubMed] [Google Scholar]
  7. Graham F. L., Smiley J., Russell W. C., Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. doi: 10.1099/0022-1317-36-1-59. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Grodzicker T., Williams J., Sharp P., Sambrook J. Physical mapping of temperature-sensitive mutations of adenoviruses. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):439–446. doi: 10.1101/sqb.1974.039.01.056. [DOI] [PubMed] [Google Scholar]
  10. Hassell J. A., Weber J. Genetic analysis of adenovirus type 2. VIII. Physical locations of temperature-sensitive mutations. J Virol. 1978 Dec;28(3):671–678. doi: 10.1128/jvi.28.3.671-678.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  12. Mulder C., Arrand J. R., Delius H., Keller W., Pettersson U., Roberts R. J., Sharp P. A. Cleavage maps of DNA from adenovirus types 2 and 5 by restriction endonucleases EcoRI and HpaI. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):397–400. doi: 10.1101/sqb.1974.039.01.051. [DOI] [PubMed] [Google Scholar]
  13. Pettersson U., Mulder C., Deluis H., Sharp P. A. Cleavage of adenovirus type 2 DNA into six unique fragments by endonuclease R-RI. Proc Natl Acad Sci U S A. 1973 Jan;70(1):200–204. doi: 10.1073/pnas.70.1.200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sambrook J., Williams J., Sharp P. A., Grodzicker T. Physical mapping of temperature-sensitive mutations of adenoviruses. J Mol Biol. 1975 Sep 25;97(3):369–390. doi: 10.1016/s0022-2836(75)80046-5. [DOI] [PubMed] [Google Scholar]
  15. Sharp P. A., Sugden B., Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. doi: 10.1021/bi00740a018. [DOI] [PubMed] [Google Scholar]
  16. Stow N. D., Wilkie N. M. An improved technique for obtaining enhanced infectivity with herpes simplex virus type 1 DNA. J Gen Virol. 1976 Dec;33(3):447–458. doi: 10.1099/0022-1317-33-3-447. [DOI] [PubMed] [Google Scholar]
  17. Takemori N. Genetic studies with tumorigenic adenoviruses. 3. Recombination in adenovirus type 12. Virology. 1972 Jan;47(1):157–167. doi: 10.1016/0042-6822(72)90249-8. [DOI] [PubMed] [Google Scholar]
  18. Takemori N., Riggs J. L., Aldrich C. Genetic studies with tumorigenic adenoviruses. I. Isolation of cytocidal (cyt) mutants of adenovirus type 12. Virology. 1968 Dec;36(4):575–586. doi: 10.1016/0042-6822(68)90189-x. [DOI] [PubMed] [Google Scholar]
  19. Willians J. F., Young C. S., Austin P. E. Genetic analysis of human adenovirus type 5 in permissive and nonpermissive cells. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):427–437. doi: 10.1101/sqb.1974.039.01.055. [DOI] [PubMed] [Google Scholar]

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

RESOURCES