Cloning and characterization of a repetitive DNA sequence specific for Trichomonas vaginalis.
A family of 650-bp-long repeats from the Trichomonas vaginalis genome, designated the Tv-E650 family, was cloned and sequenced. The nucleotide sequence is A+T-rich (73.3% A+T in the consensus sequence) and highly conserved among the 8 molecular clones analyzed. The differences among the clones are single-nucleotide and 2-nucleotide substitutions and insertions or deletions. The sequence uniformity of the clones as well as the presence of identical mutations in different clones suggest that efficient sequence homogenization mechanisms, such as gene conversion or recurring unequal crossing-over, operate in T. vaginalis. The copy number of the Tv-E650 repeats was estimated to be about 10(2)-10(3) per genome. Based on the DNA hybridization results, the Tv-E650 repeat family is conserved in all T. vaginalis strains examined, regardless of their diverse geographical origin. No hybridization of the Tv-E650 probe was found with the DNA from Trichomonas tenax, Trichomonas gallinae and Pentatrichomonas hominis, indicating that the Tv-E650 repeated sequences are species-specific. A dot blot hybridization protocol was developed which does not require isolation of DNA. By using this protocol it was possible to detect the DNA released from approximately 10(3) T. vaginalis cells per dot. These observations suggest that the Tv-E650 probe is potentially applicable to the identification and detection of T. vaginalis.
Paces
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Institute of Molecular Genetics, Czechoslovak Academy of Sciences, Prague.
Urbánková
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Urbánek
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D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000235:genetics*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D013045:Species Specificity; D014246:Trichomonas vaginalis / Q000235:genetics*
D016054:DNA, Protozoan