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RFLP analysis of PCR-amplified small subunit ribosomal DNA of three fish microsporidian species

Published online by Cambridge University Press:  01 February 2000

J. LEIRO ,
M. I. G. SISO ,
A. PARAMA ,
F. M. UBEIRA  and
M. L. SANMARTIN
J. LEIRO
Affiliation:
Laboratorio de Parasitología, Facultad de Farmacia, Universidad de Santiago de Compostela, 15706 Santiago de Compostela, Spain
M. I. G. SISO
Affiliation:
Departamento de Biología Celular y Molecular, Laboratorio de Bioquímica, Universidad de La Coruña, , Campus da Zapateira s/n, La Coruña, Spain
A. PARAMA
Affiliation:
Laboratorio de Parasitología, Facultad de Farmacia, Universidad de Santiago de Compostela, 15706 Santiago de Compostela, Spain
F. M. UBEIRA
Affiliation:
Laboratorio de Parasitología, Facultad de Farmacia, Universidad de Santiago de Compostela, 15706 Santiago de Compostela, Spain
M. L. SANMARTIN
Affiliation:
Instituto de Investigación y Análisis Alimentarios, Laboratorio de Parasitología, Facultad de Farmacia, Universidad de Santiago de Compostela, 15706 Santiago de Compostela, Spain
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Abstract

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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.

Keywords

microsporidiariboprintingphylogenysmall subunit ribosomal RNApolymerase chain reaction
Type
Research Article
Information
Parasitology , Volume 120 , Issue 2 , February 2000 , pp. 113 - 119
Copyright
2000 Cambridge University Press