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

• Published in: Fish & Shellfish Immunology
• Main Authors: Rakus, K.L., Wiegertjes, G.F., Adamek, M., Bekh, V., Stet, R.J.M., Irnazarow, I.
• Format: Article in Journal/Newspaper
• Language: English
• Published: 2008
• Subjects: atlantic salmon; disease resistance; fragment-length-polymorphism; gel-electrophoresis dgge; mhc class-ii; mutation detection; polymerase-chain-reaction; salmon salmo-salar; strand conformation polymorphism; threespine stickleback; Atlantic salmon; Salmo salar
• Online Access: https://research.wur.nl/en/publications/application-of-pcr-rf-sscp-to-study-major-histocompatibility-clas; https://doi.org/10.1016/j.fsi.2007.11.015

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