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Application of PCR-RF-SSCP to study major histocompatibility class II B polymorphism in common carp (Cyprinus carpio L.).
Rakus, Krzysztof L; Wiegertjes, Geert F; Adamek, Mikolaj; Bekh, Vitaliy; Stet, René J M; Irnazarow, Ilgiz.
Afiliación
  • Rakus KL; Polish Academy of Sciences, Institute of Ichthyobiology and Aquaculture in Golysz, Zaborze, ul. Kalinowa 2, Chybie, Poland. krzysztof.rakus@fish.edu.pl
Fish Shellfish Immunol ; 24(6): 734-44, 2008 Jun.
Article en En | MEDLINE | ID: mdl-18430588
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.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Carpas / Reacción en Cadena de la Polimerasa / Genes MHC Clase II / Polimorfismo Conformacional Retorcido-Simple Límite: Animals Idioma: En Revista: Fish Shellfish Immunol Asunto de la revista: BIOLOGIA / MEDICINA VETERINARIA Año: 2008 Tipo del documento: Article País de afiliación: Polonia Pais de publicación: Reino Unido

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Carpas / Reacción en Cadena de la Polimerasa / Genes MHC Clase II / Polimorfismo Conformacional Retorcido-Simple Límite: Animals Idioma: En Revista: Fish Shellfish Immunol Asunto de la revista: BIOLOGIA / MEDICINA VETERINARIA Año: 2008 Tipo del documento: Article País de afiliación: Polonia Pais de publicación: Reino Unido