title: Application of PCR-RF-SSCP to study major histocompatibility class II B polymorphism in common carp (Cyprinus carpio L.).
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.
authors.affiliation: Polish Academy of Sciences, Institute of Ichthyobiology and Aquaculture in Golysz, Zaborze, ul. Kalinowa 2, Chybie, Poland. krzysztof.rakus@fish.edu.pl
authors.forename: Krzysztof Ł
authors.initials: KŁ
authors.lastname: Rakus
authors.forename: Geert F
authors.initials: GF
authors.lastname: Wiegertjes
authors.forename: Mikołaj
authors.initials: M
authors.lastname: Adamek
authors.forename: Vitaliy
authors.initials: V
authors.lastname: Bekh
authors.forename: René J M
authors.initials: RJ
authors.lastname: Stet
authors.forename: Ilgiz
authors.initials: I
authors.lastname: Irnazarow
mesh_terms: D016980:Aeromonas hydrophila / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002347:Carps / Q000235:genetics* / Q000276:immunology; D005393:Fish Diseases / Q000382:microbiology; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics; D005787:Gene Frequency; D005802:Genes, MHC Class II / Q000235:genetics*; D016905:Gram-Negative Bacterial Infections / Q000662:veterinary; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D015203:Reproducibility of Results; D016415:Sequence Alignment
chemical_list: D029941:Fish Proteins