山羊DCT基因启动子区甲基化水平、SNP与毛色特征关系研究

doi:10.11843/j.issn.0366-6964.2019.02.005

Abstract

Abstract:

The purpose of this study was to investigate the effects of the methylation and SNPs in DCT gene promoter region on the coat color of goats, and to provide a theoretical basis for exploring the mechanism of DCT gene regulating the coat color of goats. Goats were used as experimental animals. The CpG island in the promoter region of goat DCT gene was predicted and two pairs of primers were designed for bisulfite methylation sequencing of the two CpG island enriched regions. Methylation level online analysis software, BISMA, was used to count the number of methylation sites and to compare the differences of methylation levels between Tangshan dairy goats(with white coat) and Nanjiang Huang goats(with black coat). Core promoter region sequence of DCT was cloned and SNPs were screened for goats with different coat colors. JASPAR and Nsite, the transcription factor online prediction software, were used to predict the transcription factor alterations with the mutation of SNPs, and promoter activity variation with the mutation of SNPs was detected. The methylation sequence and the core promoter region sequence(g.-1045--318) of goat DCT gene were successfully cloned. Six and 23 methylation sites were found in the g.-348--150 region and the g.+222-+502 region, respectively. The methylation levels of white goats at the g.+312, g.+352 and g.+400 sites and at the g.+389 and g.+404 sites were significantly and extremely significantly higher than those of black goats (P<0.05 and P<0.01), respectively. And the average methylation level of white goats in g.+222-+502 region was extremely significantly higher than that of black goats (P<0.01). The genotype compositions of the 3 SNPs in the core promoter region, g.-804 T > G, g.-705 C > T and g. -679 G > A, was different between white goats and the 3 colorful goats. SOX10 transcription factor binding site disappeared with the g.-804T > G mutation, and the promoter activity of the DCT gene was significantly decreased (P<0.05). The higher methylation level in the g.+222 -+502 region of DCT gene in white goats, and the 3 mutations at g.-804, g.-714 and g.-679 sites, especially the g.-804 T > G mutation resulted in the loss of the SOX10 transcription factor binding site and the promoter activity of DCT with G genotype was significantly decreased.In conclusion, the SNP mutation and the higher methylation level in promoter region may suppress the expression of DCT gene and form the white coat color of goat.

CLC Number:

S827.2

DU Xiaolong, WANG Qi, ZHANG Lechao, GE Linhan, LIU Xiaohui, WANG Han, LI Lanhui, LI Xianglong. Study on the Relationship between Methylation Level, SNP in Promoter Region of DCT Gene and Hair Color in Goat[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(2): 271-279.

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Study on the Relationship between Methylation Level, SNP in Promoter Region of DCT Gene and Hair Color in Goat

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