Abstract
Repetitive DNA sequences were isolated from aTheileria mutans genomic library by screening withT. mutans total DNA. DNA sequence analysis demonstrated that a section of one of the clones contained a complex series of overlapping perfect repeats ranging between 99 and 20 bp in size. TheT. mutans repetitive sequence did not contain large open reading frames (ORFs), unlikeT. parva Tpr repetitive DNA sequences. When used as a hybridisation probe the repetitive sequence revealed restriction-fragment-length polymorphisms (RFLPs) between theEcoRI-digested DNAs of twoT. mutans stocks. TheT. mutans repetitive probe gave a signal with 1 ng of purifiedT. mutans piroplasm DNA and detectedT. mutans sequences in whole-blood DNA isolated from an experimentally infected animal when the piroplasm parasitaemia was equal to or above 0.4%. Oligonucleotide primers derived from the repetitive sequence allowed more sensitive detection ofT. mutans DNA by polymerase chain reaction (PCR) amplification. Using the PCR,T. mutans DNA was amplified from an experimentally infected animal with a parasitaemia of <0.1%.
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Nucleotide sequence data reported in this paper have been submitted to the GenBank database with accession number L16682
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Bishop, R.P., Sohanpal, B.K. & Morzaria, S.P. Cloning and characterisation of a repetitive DNA sequence fromTheileria mutans: Application as a species-specific probe. Parasitol Res 80, 33–41 (1994). https://doi.org/10.1007/BF00932621
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DOI: https://doi.org/10.1007/BF00932621
Keywords
- Polymerase Chain Reaction
- Sensitive Detection
- Oligonucleotide Primer
- Repetitive Sequence
- Infected Animal