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Multiple Polymorphic Sites at the ITS and ATAN Loci in Cultured Isolates of Perkinsus marinus
Author(s) -
BROWN GWYNNE D.,
HUDSON KAREN L.,
REECE KIMBERLY S.
Publication year - 2004
Publication title -
journal of eukaryotic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 1066-5234
DOI - 10.1111/j.1550-7408.2004.tb00572.x
Subject(s) - internal transcribed spacer , biology , genetics , spacer dna , locus (genetics) , polymerase chain reaction , restriction fragment length polymorphism , gene , concerted evolution , nucleic acid sequence , genomic dna , sequence analysis , cleaved amplified polymorphic sequence , dna sequencing , microbiology and biotechnology , ribosomal rna , phylogenetics
. Sequence analysis of genomic DNA from the protozoan parasite Perkinsus marinus at two loci revealed genetic polymorphisms within and among different cultured isolates. Genomic DNA from 12 Perkinsus marinus isolates was amplified at the internal transcribed spacer region and at an anonymous locus previously identified to contain polymorphisms by restriction fragment length polymorphism analysis. Fourteen polymorphic nucleotide positions were identified at the internal transcribed spacer region; eight in internal transcribed spacer I and six in internal transcribed spacer 2. Thirteen polymorphic nucleotide sites were identified within the anonymous locus. In some instances, more than three different sequences were observed at both the internal transcribed spacer region and at the anonymous locus from a single clonal isolate, suggesting the possibility of recombination in cultured cells and/or strand jumping during the polymerase chain reaction. Intra‐isolate sequence variation (3.46% for the anonymous locus and 3.08% for internal transcribed spacer 1) was in several cases as high as inter‐isolate sequence variation, even in one isolate where recombination was not evident. High intra‐ and inter‐isolate variation detected at both loci demonstrates the importance of determining the genetic variation of each locus prior to development of sequence‐based molecular diagnostics.