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. 2014 Fall;15(4):331-5.

Polymorphism of sheep POU1F1 gene exon 6 and 3'UTR region and their association with milk production traits

O Ozmen  1 , S Kul  2 , E O Unal  3
Affiliations

Polymorphism of sheep POU1F1 gene exon 6 and 3'UTR region and their association with milk production traits

O Ozmen et al. Iran J Vet Res. 2014 Fall.

Abstract

The purpose of the study was to detect the AluI and DdeI polymorphisms within POU1F1 gene exon 6 and 3'UTR region in Turkish sheep breeds, and their association with milk productive traits. Jugular blood samples were collected from 50 Sakiz, 50 White Karaman, and 50 Awassi sheep using EDTA as an anticoagulant. PCR-RFLP and sequencing analysis were performed to investigate possible polymorphisms in the exon 6 and 3' flanking region of the sheep POU1F1 gene. The PCR products were digested with restriction endonuclease AluI and DdeI, and biallelic polymorphism was found with restriction endonuclease AluI and two genotypes (TT (296 bp and 173 bp) and CC (235 bp, 173 bp and 61 bp)) were detected. White Karaman and Awassi breeds did not show polymorphisms for AluI restriction sites. No polymorphism at the DdeI cleavage sites was detected in the three sheep breeds. Significant statistical results were found in milk yield ((***)P0.001), fat ((***)P0.001) and lactose ((*)P0.05) values with TT and CC genotypes, however no significant association of TT and CC genotypes with protein values was detected (P>0.05) and individuals with genotype TT had a superior milk yield in Sakiz sheep breeds. As sequence results, seven variation points were determined for exon 6 (g.185T>C) and 3'UTR (g.220G>A, g.229C>T, g.248C>T, g.250A>T, g.255T>C, g.258C>T) of the sheep POU1F1 gene. We have reported here for the first time single nucleotide polymorphisms of the POU1F1 gene for both exon 6 and 3'UTR and its effects on milk traits in Turkish sheep breeds were evaluated.

Keywords: Milk traits; POU1F1 gene; Polymorphism; Turkish sheep breeds.

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Figures

Fig. 1
Fig. 1
Agarose gel electrophoresis band patterns after digestion with AluI endonuclease within the exon 6 and its flanking region sequence of the sheep POU1F1 gene. Lane M: 100 bp DNA marker. Lane 2-4: TT genotype (296 bp and 173 bp), and Lane 5-9: CC genotype (235 bp, 173 bp and 61 bp). As one small band (61 bp) was invisible on 4% agarose gel only two bands were visible for CC genotype
Fig. 2
Fig. 2
The structure of sheep POU1F1 gene and SNPs location for exon 6 and 3'UTR and AluI, DdeI enzyme restriction sites. Black dots (●) represent mutation points
Fig. 3
Fig. 3
DNA sequence of genotypes TT at the 236 locus and detected variation points in sheep POU1F1 gene exon 6 and 3'-UTR region
Fig. 4
Fig. 4
Agarose gel electrophoresis band patterns after digestion with DdeI endonuclease within the exon 6 and its flanking region sequence of the sheep POU1F1 gene. Lane M: 100 bp DNA marker. Lane 2-7: D1D1 genotype (199 bp, 118 bp, 83 bp and 58 bp). As one small band (11 bp) was invisible on 4% agarose gel only four bands were visible for CC genotype
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