Molecular Biotechnology

, Volume 23, Issue 1, pp 83–86 | Cite as

Universal primers for the PCR-mediated amplification of DNA 1: A satellite-like molecule associated with begomovirus-DNA β complexes

Hints And Tips


DNA 1 is a single-stranded DNA molecule of approximately 1370 nucleotides. It is associated with monopartite geminiviruses of the genus Begomovirus, which require a DNA β component for symptomatic infection. The DNA 1 molecule requires the helper begomovirus for movement in plants, but is capable of self-replication. We designed two abutting primer pairs (DNA101/DNA102 and UN101/UN102) to conserved sequences of DNA 1. This allowed polymerase chain reaction-mediated amplification of the full-length molecule from total nucleic acid extracts produced from various host plants from geographically distinct, worldwide locations. These primers are useful both as diagnostic probes and for producing full-length infectious clones for in planta studies.

Index Entries

DNA 1 begomovirus PCR oligonucleotide primers DNA β 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Lazarowitz, S. G. (1992) Geminiviruses: genome structure and gene function. Crit. Rev. Plant Sci. 11, 327–349.Google Scholar
  2. 2.
    Rybicki, E. P., Briddon, R. W., Brown, et al. (2000) Geminiviridae. In Virus Taxonomy. Seventh Report of the International Committee on Taxonomy of Viruses (van Regenmortel, M. H. V., Fauquet, C. M., Bishop, D. H. L., et al. eds.), Academic Press, San Diego, pp. 285–297.Google Scholar
  3. 3.
    Czosnek, H. and Laterrot, H. (1997) A worldwide survey of tomato yellow leaf curl viruses. Arch. Virol. 142, 1391–1406.PubMedCrossRefGoogle Scholar
  4. 4.
    Briddon, R., Mansoor, S., Bedford, I., et al. (2001) Identification of DNA components required for induction of cotton leaf curl disease. Virology 285, 234–243.PubMedCrossRefGoogle Scholar
  5. 5.
    Saunders, K., Bedford, I. D., Briddon, R. W., Markham, P. G., Wong, S. M., and Stanley, J. (2000) A novel virus complex causes Ageratum yellow vein disease. Proc. Natl. Acad. Sci. USA 97, 6890–6895.PubMedCrossRefGoogle Scholar
  6. 6.
    Mansoor, S., Khan, S. H., Bashir, A., et al. (1999) Identification of a novel circular single-stranded DNA associated with cotton leaf curl disease in Pakistan. Virology 259, 190–199.PubMedCrossRefGoogle Scholar
  7. 7.
    Saunders, K. and Stanley, J. (1999) A nanovirus-like component associated with yellow vein disease of Ageratum yellow vein disease of Ageratum conyzoides: evidence for interfamilial recombination between plant disease plant DNA viruses. Virology 264, 142–152.PubMedCrossRefGoogle Scholar
  8. 8.
    Briddon, R. W., Bull, S. E., Mansoor, S., Amin, I., and Markham, P. G. (2002) Universal primers for the PCR-mediated amplification of DNA β; a molecule associated with some monopartite begomoviruses. Mol. Biotech. 20, 315–318.CrossRefGoogle Scholar
  9. 9.
    Briddon, R. W. and Markham, P. G. (1994) Universal primers for the PCR amplification of dicot-infecting geminiviruses. Mol. Biotech. 1, 202–205.CrossRefGoogle Scholar
  10. 10.
    Rojas, M. R., Gilbertson, R. L., Russell, D. R., and Maxwell, D. P. (1993) Use of degenerate primers in the polymerase chain reaction to detect whitefly-transmitted geminiviruses. Plant Dis. 77, 340–347.CrossRefGoogle Scholar
  11. 11.
    Doyle, J. J. and Doyle, J. L. (1987) A rapid DNA isolation procedure for small quantities of fresh leaf material. Phytochem. Bull. 19, 11–15.Google Scholar
  12. 12.
    Covey, S. N. and Hull, R. (1981) Transcription of cauliflower mosaic virus DNA. Detection of transcripts, properties and location of the gene encoding the virus inclusion body protein. Virology 111, 463–474.CrossRefPubMedGoogle Scholar
  13. 13.
    Saunders, K., Bedford, I. D., and Stanley, J. (2001) Pathogenicity of a natural recombinant associated with Ageratum yellow vein disease: implications for geminivirus evolution and disease aetiology. Virology 282, 38–47.PubMedCrossRefGoogle Scholar
  14. 14.
    Stanley, J., Saunders, K., Pinner, M. S., and Wong, S. M. (1997) Novel defective interfering DNAs associated with Ageratum yellow vein geminivirus infection of Ageratum conyzoides. Virology 239, 87–96.PubMedCrossRefGoogle Scholar
  15. 15.
    Liu, Y., Robinson, D. J., and Harrison, B. D. (1998) Defective forms of cotton leaf curl virus DNA-A that have different combinations of sequence deletion, duplication, inversion and rearrangement. J. Gen. Virol. 79, 1501–1508.PubMedGoogle Scholar

Copyright information

© Humana Press Inc 2003

Authors and Affiliations

  1. 1.Department of Disease and Stress BiologyJohn Innes CentreNorwichUK

Personalised recommendations