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

Parasitology. 2000 Feb:120 ( Pt 2):113-9. doi: 10.1017/s0031182099005405.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA Primers / chemistry
  • DNA, Protozoan / chemistry
  • DNA, Protozoan / isolation & purification
  • DNA, Ribosomal / chemistry*
  • DNA, Ribosomal / isolation & purification
  • Deoxyribonucleases, Type II Site-Specific / chemistry
  • Electrophoresis, Agar Gel / veterinary
  • Fishes / parasitology*
  • Heteroduplex Analysis / veterinary
  • Image Processing, Computer-Assisted
  • Microsporida / classification
  • Microsporida / genetics*
  • Microsporidiosis / veterinary*
  • Phylogeny
  • Polymerase Chain Reaction / veterinary
  • Polymorphism, Restriction Fragment Length

Substances

  • DNA Primers
  • DNA, Protozoan
  • DNA, Ribosomal
  • endodeoxyribonuclease AluI
  • endodeoxyribonuclease DdeI
  • Deoxyribonucleases, Type II Site-Specific