Genotyping pigs of Ukrainian breeds by microsatellite loci with a tetranucleotide motif

Abstract

The aim of the research was genotyping animals of Ukrainian pig breeds according to microsatellite loci of the genome for further creation and implementation of the methodology for determining gene complexes of pig adaptability. Research methods: zootechnical – the analysis of genealogy and the productivity indexes of animals of different breeds; bioinformation - work with databases of the primary genome sequences for the design of oligonucleotide primers; molecular genetic - nucleic acid extraction, the determination of purity and concentration of DNA preparations, the amplification of microsatellite loci in PCR with subsequent electrophoretic separation of the amplification products in denaturing gel; statistical – the determination of population-genetic parameters on the basis of typing data on microsatellite loci, the relationship of microsatellite loci with performance indexes, the search for adaptive complexes. It was carried out genotyping animals of the Ukrainian meat breed in SE “RF Elita MIW named after V. M. Remeslo NAAS "of Kyiv region for 6 microsatellite loci with tetranucleotide repeats, a panel of primers of its own design. It was found the fact that the average number of alleles per locus was 5.2 alleles. The level of actual heterozygosity was 0.830, while the expected heterozygosity was 0.780. However, this difference is not significant. DNA typing animals for 6 loci of microsatellite DNA of 17 heads of Poltava meat breed of SE "EB Nadiia" was carried out. As a result of genotyping, the average number of alleles per locus was 3.8. The actual and expected heterozygosity levels were 0.680 and 0.730, respectively. By loci of microsatellite DNA, we found in animals of Myrhorod breed: for locus FH3628 – 5 alleles of 200-228 nucleotides, FH1865 – 5 alleles (size from 109 to 129 nucleotides), FH4219 – 2 alleles of 80 and 92 nucleotides, FH1885 – 4 alleles from 205 up to 213 nucleotides, FH3764 – 2 alleles 138 and 146 nucleotides, and for FH4231 – 2 alleles 116 and 120 nucleotides. The level of actual heterozygosity was 0.860, while the expected heterozygosity was 0.810. Thus, in three breeds of Ukrainian selection for 6 loci of microsatellite DNA with tetranucleotide repeat, the level of actual heterozygosity was in the range of 0.680 in animals of Poltava meat breed to 0.860 in animals of Myrhorod breed. The level of expected heterozygosity is 0.730 in animals of Poltava meat breed to 0.810 in Myrhorod breed. The high level of heterozygosity, as well as the average number of alleles per locus allows to search for alleles and genotypes associated with the adaptability of animals in breeds of pigs of Ukrainian selection. Key words: pigs, microsatellite loci, genome, average number of alleles, level of heterozygosity, population, variability.