| The polymorphisms of13microsatellite loci of116Gansu Alpine Merino sheep wasdetected using multiplex polymerase chain reaction and DNA sequencing technology, andthe paternity testing was studied combined with the parents of this record. The purpose isto supply effective microsatellite locus for germplasm resourses evaluation and paternitytesting of sheep. To investigate polymorphism, the number of alleles, single nucleotidePolymorphisms (SNPs) sites, variation type, the genetic relationship and evolutionarysignificance of the alleles of the sheep MHC (Ovar) class II DRB1locus, we amplifiedOvar-DRB1exon3by polymerase chain reaction-single-strand conformationpolymorphism (PCR-SSCP). Results demonstrated that all above results suggested thatthere were a relatively high genetic diversity in Gansu Alpine Merino.13microsatelliteloci altogether discovered111alleles. The number of alleles was most inDRB1-INTRO2(14), and the least in SRCRCP5(5). The heterozygosity was highest inILSTS011(0.888), while the lowest was MAF214(0.500). The mean observedheterozygosity, the expected heterozygosity, and the polymorphism information content(PIC) were0.743,0.727, and0.689, respectively. The calculate exclusion probability (CPE)of the13locus was0.99994. The CPE of the11locus which were highly polymorphicwere0.99985, and12locus were0.99991. MAF214had a smaller effect on CPE andDRB1-INTRO2had a higher effect on CPE. In the study of the sheep named Gansu AlpineMerino, there were identified1previously published alleles,5new alleles, and11nucleotide polymorphism sites. The phylogenetic tree results showed that the6haplotypesof Ovar-DRB1exon3could divide into two clusters, and allele A was the most commonallele. The results suggest that Ovar-DRB1exon3has high level of sequencepolymorphism(PIC>0.5), and it is differentiated into two major category alleles from twomutant alleles at first. The genotypes of Ovar-DRB1exon3unfitted with Hardy-Weinbergequilibrium in the detection region. |
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