engleski

Comparison of the PCR methods for assessing variability of Major Histocompatibility Complex class II DRB1 locus in wild boar (Sus scrofa L.) from Croatia

Major histocompatibility complex genes encode cell-surface glycoproteins that present antigens to T cells, a process essential for triggering an appropriate immune response. MHC contains some of the most variable genes in vertebrates whose diversity has been driven by evolutionary processes over long period of time. Their variability reflects the ability of a population to confront various pathogens and strong association between MHC variability and resistance to infectious organisms has been established. Wild boar's widespread distribution and potential to host many parasite species makes it suitable for testing that correlation. The goal of this study was to improve our method for assessing variability of the MHC class II DRB1 locus, since our preliminary analysis revealed only 5.5% of heterozygous individuals, which strongly indicated existence of null-alleles. In this study we compare results obtained by the two PCR methods, differing in primer sequences. We analysed 155 samples of wild boars from Croatia. Muscle samples were collected following regular hunting operations and DNA was extracted using commercial kit. Polymerase chain reaction was used to amplify the exon 2 of DRB1 locus with different primers than in our previous preliminary research. Furthermore, the primers used in this research enabled amplification of the whole exon 2 (267 bp), while previous method amplified 198 bp of the exon 2. We successfully amplified and sequenced 124 individuals of which 73.4% were heterozygotes. We identified 19 different DRB1 alleles, seven of which were new, while only 11 alleles were identified in our previous research. In conclusion, the PCR method with different primer set proved to be more efficient for identification of all/majority of alleles present in a population of wild boar from Croatia.

wild boar ; MHC genes ; variability ; comparison of methods

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