Archives of Virology

, Volume 124, Issue 3–4, pp 301–319 | Cite as

Characterization of viral DNAs from cells infected with chicken anaemia agent: sequence analysis of the cloned replicative form and transfection capabilities of cloned genome fragments

  • B. M. Meehan
  • D. Todd
  • J. L. Creelan
  • J. A. P. Earle
  • E. M. Hoey
  • M. S. McNulty
Original Papers


The viral DNAs induced by the unclassified animal virus, chicken anaemia agent (CAA), during replication in MDCC-MSB 1 cells have been investigated. Analyses after S1 nuclease, restriction endonuclease and denaturation treatments indicated that infected cell extracts contained genome-size, single-stranded DNA (Mr 2.3 kb), closed and open circular, double-stranded replicative form (RF) DNAs (Mr 2.3 kbp) and a population of smaller doublestranded DNAs (Mr 0.8 kbp). Recombinant plasmids containing 2.3 kbp CAA RF fragments cloned at the PstI, BamHI and EcoRI sites failed to transfect MDCC-MSB 1 cells. However, one plasmid, which contained two 2.3 kbp CAA RF fragments ligated in tandem at the PstI site, and cloned 2.3 kbp PstI, BamHI and EcoRI fragments, excised from their respective plasmids by restriction endonuclease digestion, were capable of transfection.

The nucleotide sequence of the circular genome (2298 bp) of the Cux-1 isolate of CAA has indicated the presence of three overlapping open reading frames (ORFs) of 52 kDa, 24 kDa and 13 kDa on one strand. The existence of these ORFs was corroborated by analyses of partial sequences from three other isolates. The non-coding region of the CAA genome contained sequences with putative regulatory function. These results are discussed in relation to the “rolling circle” model of DNA replication.


Viral DNAs Restriction Endonuclease Digestion Genome Fragment EcoRI Site EcoRI Fragment 
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  1. 1.
    Briggs MR, Kadonaga JT, Bell SP, Tjian R (1986) Purification and biochemical characterization of the promoter-specific transcription factor, Sp 1. Science 234: 47–52Google Scholar
  2. 2.
    Claessens JAJ, Schrier CC, Mockett APA, Jagt EHJM, Sondermeijer PAJ (1991) Molecular cloning and sequence analysis of the genome of chicken anaemia agent. J Gen Virol 72: 2003–2006Google Scholar
  3. 3.
    Connor TJ, McNeilly F, Firth GA, McNulty MS (1991) Biological characterisation of Australian isolates of chicken anaemia agent. Aust Vet J 68: 199–201Google Scholar
  4. 4.
    Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the Vax. Nucleic Acids Res 12: 387–395Google Scholar
  5. 5.
    Feinberg A, Vogelstein B (1984) A technique for radio-labelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 137: 266–267Google Scholar
  6. 6.
    Fisch TM, Prywes R, Roeder RG (1987)c-fos sequences necessary for basal expression and induction by epidermal growth factor, 12-O-tetradecanoyl phorbol-13-acetate, and the calcium ionophore. Mol Cell Biol 7: 3490–3502Google Scholar
  7. 7.
    Gelderblom G, Kling S, Lurz R, Tischer I, Von Bulow V (1989) Morphological characterisation of chicken anaemia agent (CAA). Arch Virol 109: 115–120Google Scholar
  8. 8.
    Goryo M, Suwa T, Matsumoto S, Umemura T, Itakura C (1987) Serial propagation and purification of chicken anaemia agent in MDCC-MSB 1 cell line. Avian Pathol 16: 149–163Google Scholar
  9. 9.
    Hirt B (1967) Selective extraction of polyoma virus DNA from infected cell cultures. J Mol Biol 36: 365–369Google Scholar
  10. 10.
    Jones KA, Kadonaga JT, Rosenfeld PJ, Kelly TJ, Tjian R (1987) A cellular DNA-binding protein that activates eukaryotic transcription and DNA replication. Cell 48: 79–89Google Scholar
  11. 11.
    Kornberg A (1980, Supplement 1982) DNA replication. Freeman, San FranciscoGoogle Scholar
  12. 12.
    Kozak M (1987) An analysis of 5′-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res 15: 8125–8148Google Scholar
  13. 13.
    Leung K, Nabel GJ (1988) HTLV-1 transactivator induces interleukin-2 receptor expression through an NF-kB-like factor. Nature 333: 776–778Google Scholar
  14. 14.
    Lewis CD, Clark SP, Felsenfeld G, Gould H (1988) An erythrocyte-specific protein that binds to the poly (dG) region of the chicken β-globin gene promoter. Genes Dev 2: 863–873Google Scholar
  15. 15.
    Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, New YorkGoogle Scholar
  16. 16.
    Mankertz J, Buhk H-J (1990) Abstract, VIII international congress of virology, p 380Google Scholar
  17. 17.
    Matsui T, Segall J, Weil PA, Roeder RG (1980) Multiple factors for accurate initiation of transcription by purified RNA polymerase II. Biol Chem 225: 11992–11996Google Scholar
  18. 18.
    McNulty MS, Connor TJ, McNeilly F, Kirkpatrick KS, McFerran JB (1988) A serological survey of domestic poultry in the United Kingdom for antibody to chicken anaemia agent. Avian Pathol 17: 315–324Google Scholar
  19. 19.
    McNulty MS, Connor TJ, McNeilly F, McLoughlin MF, Kirkpatrick KS (1990) Preliminary characterisation of isolates of chicken anaemia agent from the United Kingdom. Avian Pathol 19: 67–93Google Scholar
  20. 20.
    McNulty MS, Connor TJ, McNeilly F, Spackman D (1989) Chicken anemia agent in the United States: isolation of the virus and detection of antibody in broiler breeder flocks. Avian Dis 33: 691–694Google Scholar
  21. 21.
    McNulty MS, Curran WL, Todd D, Mackie DP (1990) Chicken anemia agent: an electron microscopic study. Avian Dis 34: 736–743Google Scholar
  22. 22.
    Messing J, Vieira T (1982) A new pair of M 13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene 19: 269–276Google Scholar
  23. 23.
    Molitor TW, Joo HS, Collet MS (1984) Porcine parvovirus DNA: characterization of the genomic and replicative form DNA of two virus isolates. Virology 137: 241–254Google Scholar
  24. 24.
    Nevins JR (1983) The pathway of eukaryotic mRNA formation. Annu Rev Biochem 52: 441–446Google Scholar
  25. 25.
    Noteborn MHM, de Boer GF, van Roozelaar DJ, Katreman C, Kranenburg O, Vos JG, Jeurissen SHM, Hoeben RC, Zantema A, Koch G, van Ormondt H, van der Elb AJ (1991) Characterization of cloned chicken anemia virus DNA that contains all elements for the infectious replication cycle. J Virol 65: 3131–3139Google Scholar
  26. 26.
    Pettersson M, Schaffner W (1987) A purine-rich DNA sequence motif present in SV 40 and lymphotrophic papovaviruses binds a lymphoid-specific factor and contributes to enhancer activity in lymphoid cells. Genes Dev 1: 962–972Google Scholar
  27. 27.
    Plumb M, Frampton J, Wainwright H, Walker M, Macleod K, Goodwin G, Harrison P (1989) GATAAG; Acis-control region binding an erythroid-specific nuclear factor with a role in globin and non-globin gene expression. Nucleic Acids 17: 73–92Google Scholar
  28. 28.
    Ritchie BW, Niagro FD, Lukert PD, Steffens WL, Latimer KS (1989) Characterization of a new virus from cockatoos with psittacine beak and feather disease. Virology 171: 83–88Google Scholar
  29. 29.
    Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467Google Scholar
  30. 30.
    Sassone-Corsi P (1988) Cyclic AMP induction of early adenovirus promoters involves sequences required for EIA trans-activation. Proc Natl Acad Sci USA 85: 7192–7196Google Scholar
  31. 31.
    Sompayrac LM, Danna KJ (1981) Efficient infection of monkey cells with DNA of simian virus 40. Proc Natl Acad Sci USA 78: 7575–7578Google Scholar
  32. 32.
    Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503–517Google Scholar
  33. 33.
    Tischer I, Buhk H-J (1988) Viral DNA from cells infected with porcine circovirus. Zentralbl Bakteriol Mikrobiol Hyg [A] 270: 280–287Google Scholar
  34. 34.
    Tischer I, Gelderblom H, Vetterman W, Koch MA (1982) A very small porcine virus with circular, single-stranded DNA. Nature 295: 64–66Google Scholar
  35. 35.
    Todd D, Creelan JL, Mackie DP, Rixon F, McNulty MS (1990) Purification and biochemical characterization of chicken anaemia agent. J Gen Virol 71: 819–823Google Scholar
  36. 36.
    Todd D, Creelan JL, McNulty MS (1991) Dot blot hybridization assay for chicken anemia agent using a cloned DNA probe. J Clin Microbiol 29: 933–939Google Scholar
  37. 37.
    Todd D, Niagro FD, Ritchie BW, Curran W, Allan GM, Lukert PD, Latimer KS, Steffens WL, McNulty MS (1991) Comparison of three animal viruses with circular, single-stranded DNA genomes. Arch Virol 116: 129–135Google Scholar
  38. 38.
    Yano O, Kanellopoulos J, Kieran M, Le Bail O, Israel A, Kourilsky P (1987) Purification of KBF 1, a common factor binding both H-2 and β 2-microglobulin enhancers. EMBO J 6: 3317–3324Google Scholar
  39. 39.
    Yuasa N, Tangiguchi T, Yoshshida I (1979) Isolation and some properties of an agent inducing anaemia in chicks. Avian Dis 23: 366–385Google Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • B. M. Meehan
    • 1
  • D. Todd
    • 1
  • J. L. Creelan
    • 1
  • J. A. P. Earle
    • 2
  • E. M. Hoey
    • 2
  • M. S. McNulty
    • 1
  1. 1.Department of Agriculture for Northern IrelandVeterinary Sciences DivisionBelfast
  2. 2.Division of Genetic EngineeringThe Queen's University of BelfastBelfastUK

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