RFLP analysis of PCR-amplified small subunit ribosomal DNA of three fish microsporidian species.
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.
Leiro
J
J
Laboratorio de Parasitología, Facultad de Farmacia, Universidad de Santiago de Compostela, Spain. mpleiro@usc.es
Siso
M I
MI
Parama
A
A
Ubeira
F M
FM
Sanmartin
M L
ML
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
D017931:DNA Primers; D016054:DNA, Protozoan; D004275:DNA, Ribosomal; C024669:endodeoxyribonuclease AluI; C027908:endodeoxyribonuclease DdeI; D015252:Deoxyribonucleases, Type II Site-Specific