Abstract
The random amplification of polymorphic DNA (RAPD method), was used to derive fragments of DNA of the coccidial parasiteEimeria tenella for use both as genetic markers and as an aid for the discrimination of different wild-type and attenuated populations. Of 41 arbitrarily chosen 9-to 27-mer primers, 24 yielded useful arrays of fragments following low-stringency annealing conditions and the resultant profiles were generally very reproducible. One non-variant fragment of 2 kb hybridised to a single chromosome (number 12) and four variant fragments were identified. These results strongly suggest that the RAPD method may be an extremely useful tool for studies on various aspects of the genetic organisation of coccidial parasites.
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References
Akopyanz N, Bukanov NO, Kresovich S, Berg DE (1992) DNA diversity among clinical isolates ofHelicobacter pylori detected by PCR-based RAPD fingerprinting. Nucleic Acids Res 20:5137–5142
Barker D, Schafer M, White R (1984) Restriction sites containing CpG show a higher frequency of polymorphism in human DNA. Cell 36:131–138
Barral V, This P, Imbert-Establet D, Combes C, Delseny M (1993) Genetic variability and evolution of the schistosome genome analysed by using random amplified polymorphic DNA markers. Mol Biochem Parasitol 59:211–222
Jeffers TK (1975) Attenuation ofEimeria tenella through selection for precociousness. J Parasitol 61:1083–1090
Long PL (1972)Eimeria tenella: reproduction, pathogenicity and immunogenicity of a strain maintained in chicken embryos by serial passage. J Comp Pathol 82:439–445
MacPherson JM, Gajadhar AA (1993) Differentiation of sevenEimeria species by random amplified polymorphic DNA. Vet Parasitol 45:257–266
McDonald V, Shirley MW, Millard BJ (1986) A comparative study of two lines ofEimeria tenella attenuated either by selection for precocious development in the chicken or by growth in chicken embryos. Avian Pathol 15:323–335
McDougald LR, Jeffers TK (1976) Comparative in vitro development of precocious and normal strains ofEimeria tenella. J Protozool 23:530–534
Neto ED, Pereira de Souza C, Rollinson D, Katz N, Pena SDJ, Simpson AJG (1993) The random amplification of polymorphic DNA allows the identification of strains and species of schistosome. Mol Biochem Parasitol 57:83–88
Procunier JD, Fernando MA, Barta JR (1993) Species and strain differentiation ofEimeria spp. of the domestic fowl using DNA polymorphisms amplified by arbitrary primers. Parasitol Res 79:98–102
Sambrook J, Fritsch EF Maniatis T (1989) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Schmatz DM, Crane MSJ, Murray PK (1984) Purification ofEimeria sporozoites by DE-52 anion exchange chromotography. J Protozool 31:181–183
Shirley MW (1975) Enzyme variation inEimeria species of the chicken. Parasitology 71:369–376
Shirley MW (1986) Research in avian coccidiosis. In: McDougald LR, Joyner LP, Long PL (eds) Proceedings of the Georgia Coccidiosis Conference. University of Georgia Press, Athens, Georgia, pp 13–35
Shirley MW (1994) Coccidial parasites from the chicken: discrimination of different populations ofEimeria tenella by DNA hybridisation. Res Vet Sci (in press)
Shirley MW, Millard BJ (1976), Some observations on the sexual differentiation ofEimeria tenella using single sporozoite infections in chicken embryos. Parasitology 73:337–341
Shirley MW, Chapman HD, Jeffers TK, Bedrnik P (1989) Enzyme variation and pathogenicity of recent field isolates ofEimeria tenella. Res Vet Sci 46:79–83
Shirley MW, Kemp DJ, Pallister J, Prowse SJ (1990) A molecular karyotype ofEimeria tenella as revealed by contour-clamped homogeneous electric field electrophoresis. Mol Biochem Parasitol 38:169–174
Waitumbi JN, Murphy NB (1993) Inter-and intra-specific differentiation of trypanosomes by genomic fingerprinting, with arbitrary primers. Mol Biochem Parasitol, 58:181–186
Welsh J, McClelland M (1990) Fingerprinting genomes with PCR with arbitrary primers. Nucleic Acids Res 18:7213–7218
Welsh J, McClelland M (1991) Genomic fingerprinting, using arbitrarily primed PCR and a matrix of pairwise combinations of primers. Nucleic Acids Res 19:5275–5279
Williams JGK, Kubelik, AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535
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Shirley, M.W., Bumstead, N. Intra-specific variation withinEimeria tenella detected by the random amplification of polymorphic DNA. Parasitol Res 80, 346–351 (1994). https://doi.org/10.1007/BF02351878
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DOI: https://doi.org/10.1007/BF02351878