title: RFLP analysis of PCR-amplified small subunit ribosomal DNA of three fish microsporidian species.
abstract: The phylogenetic relationships of the microsporidian species Microgemma caulleryi, Pleistophora finisterrensis and Tetramicra brevifilum were investigated on the basis of restriction fragment length polymorphism (RFLP) analysis of PCR-amplified small-subunit rDNA (SSUrDNA). Using PCR primers specific for microsporidian SSUrDNA, a single product was obtained from each species, and heteroduplex analysis indicated a high degree of sequence homology among the 3 products. In RFLP analysis of the PCR-amplified SSUrDNA, the enzymes AluI and DdeI gave restriction patterns that differed among all 3 species. Phylogenetic analysis using restriction patterns as differential characters indicated that Microgemma caulleryi and Tetramicra brevifilum are more closely related to each other than to Pleistophora finisterrensis.
authors.affiliation: Laboratorio de Parasitología, Facultad de Farmacia, Universidad de Santiago de Compostela, Spain. mpleiro@usc.es
authors.forename: J
authors.initials: J
authors.lastname: Leiro
authors.forename: M I
authors.initials: MI
authors.lastname: Siso
authors.forename: A
authors.initials: A
authors.lastname: Parama
authors.forename: F M
authors.initials: FM
authors.lastname: Ubeira
authors.forename: M L
authors.initials: ML
authors.lastname: Sanmartin
mesh_terms: D000818:Animals; D017931:DNA Primers / Q000737:chemistry; D016054:DNA, Protozoan / Q000737:chemistry / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000737:chemistry* / Q000302:isolation & purification; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry; D004587:Electrophoresis, Agar Gel / Q000662:veterinary; D005399:Fishes / Q000469:parasitology*; D020180:Heteroduplex Analysis / Q000662:veterinary; D007091:Image Processing, Computer-Assisted; D016816:Microsporida / Q000145:classification / Q000235:genetics*; D016881:Microsporidiosis / Q000662:veterinary*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D017931:DNA Primers; D016054:DNA, Protozoan; D004275:DNA, Ribosomal; C024669:endodeoxyribonuclease AluI; C027908:endodeoxyribonuclease DdeI; D015252:Deoxyribonucleases, Type II Site-Specific