Application of PCR-RF-SSCP to study major histocompatibility class II B polymorphism in common carp (Cyprinus carpio L.)

K.L. Rakus, G.F. Wiegertjes, M. Adamek, V. Bekh, R.J.M. Stet, I. Irnazarow

Research output: Contribution to journalArticleAcademicpeer-review

22 Citations (Scopus)

Abstract

A variety of methods have been applied for the characterization of major histocompatibility (MH) polymorphism in fish. We optimized a technique designated polymerase chain reaction-restriction fragments-single strand conformation polymorphism (PCR-RF-SSCP) for screening a large number of individuals for the Cyca-DAB1 and Cyca-DAB2 genes polymorphism in common carp. The advantages of this technique are simplicity, high sensitivity and low costs. PCR-RF-SSCP analysis revealed different genotypes consisting of unique combinations of the Cyca-DAB1 and Cyca-DAB2 sequences with the number of SSCP bands clearly correlating with the degree of heterozygosity of the Cyca-DAB1 and Cyca-DAB2 genes. We found four alleles for Cyca-DAB1 (*02-*05) gene but only one allele for Cyca-DAB2 (*02) and noted that the Cyca-DAB2 gene was either homozygous or absent. PCR-RF-SSCP analysis of n=79 carp individuals challenged with Aeromonas hydrophila indicated that individuals bearing no Cyca-DAB2 gene showed higher cumulative mortality and lower bacterial agglutination titers during the experiment. We suggest that our PCR-RF-SSCP method can be used to study correlations of different MH class II B genotypes/alleles with resistance of common carp to specific pathogens
Original languageEnglish
Pages (from-to)734-744
JournalFish and Shellfish Immunology
Volume24
Issue number6
DOIs
Publication statusPublished - 2008

Keywords

  • mhc class-ii
  • salmon salmo-salar
  • strand conformation polymorphism
  • polymerase-chain-reaction
  • fragment-length-polymorphism
  • gel-electrophoresis dgge
  • atlantic salmon
  • threespine stickleback
  • disease resistance
  • mutation detection

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