Do variability and expression level of fish MHC genes play crucial role in parasite load and host immunocompetence?
| Authors | |
|---|---|
| Year of publication | 2014 |
| Type | Article in Proceedings |
| Conference | 3rd Workshop of European Centre of Ichthyoparasitology |
| MU Faculty or unit | |
| Citation | |
| Field | Genetics and molecular biology |
| Keywords | MHC; gene expression; HRM; Cyprinidae |
| Description | We have confirmed extremely high level of allelic diversity in several fish genera by classical genotyping. We used two basic approaches for this purpose. First, the older, was CE-SSCP (Capillary Electrophoresis-Single Stranded Conformational Polymorphism) of PCR amplicon clones combined with automatic sequencing based on Sanger technology (ABI 3130 Genetic Analyzer). The second, modern, was 454 clonal pyrosequencing of heterozygous amplicons (454 GS FLX platform with Titanium chemistry). Recently, using CFX96 real time PCR system, we test also the expression levels of fish MHC genes (DAB1 and DAB3) by quantitative real-time PCR (qPCR) and we launched a new approach for MHC genes variability tests - high resolution melting analysis (HRM), too. After completing "HRM allele profile database" and finish analyses of qPCR we will be exploring associations with parasite load in chosen Cyprinid species. |
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