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

Fish Shellfish Immunol. 2008 Jun;24(6):734-44. doi: 10.1016/j.fsi.2007.11.015. Epub 2007 Dec 5.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aeromonas hydrophila / physiology
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Carps / genetics*
  • Carps / immunology
  • Fish Diseases / microbiology
  • Fish Proteins / chemistry
  • Fish Proteins / genetics
  • Gene Frequency
  • Genes, MHC Class II / genetics*
  • Gram-Negative Bacterial Infections / veterinary
  • Molecular Sequence Data
  • Phylogeny
  • Polymerase Chain Reaction*
  • Polymorphism, Single-Stranded Conformational*
  • Reproducibility of Results
  • Sequence Alignment

Substances

  • Fish Proteins

Associated data

  • GENBANK/EU203666
  • GENBANK/EU203667
  • GENBANK/EU203668
  • GENBANK/EU203669
  • GENBANK/EU203670