UCP3基因在巴马猪和藏猪脂肪组织中的表达和甲基化分析

doi:10.16431/j.cnki.1671-7236.2019.12.020

Abstract

Abstract: The aim of this study was to investigate the expression and methylation levels of the uncoupling protein 3 (UCP3) in subcutaneous adipose tissue of Bama and Tibetan pigs.The expression of UCP3 gene at mRNA level was detected by Real-time quantitative PCR method.CpG islands were predicted for the sequence in pig UCP3 gene promoter region (-3 580 to +920 bp) using online software MethPrimer.The difference methylation level of UCP3 gene in Bama and Tibetan pigs was detected by bisulfite sequencing PCR (BSP).The results showed that the expression of UCP3 gene in subcutaneous adipose tissue of Bama pigs was significantly higher than that of Tibetan pigs (P<0.05).Three CpG methylation islands were predicted in the promoter region of UCP3 gene,which were recorded as CpG island1 (-3 171 to -2 928 bp),CpG island2 (-154 to -2 bp) and CpG island3 (+648 to +806 bp).The methylation levels of CpG island1 and CpG island3 were less different in Bama and Tibetan pigs,while the methylation level of CpG island2 (42.61%) was significantly higher in Tibetan pigs than that in Bama pigs (24.49%).A black and white dot maps were built based on the CpG island2 methylation of Bama and Tibetan pigs,the methylation frequency of CpG sites 4,8,9,10,11,12 and 15 in Tibetan pigs were 28.26%,17.39%,26.09%,26.09%,26.09%,23.91% and 34.78% higher than that in Bama pigs,respectively.In addition,three transcription factor binding sites (SP2,PPARγ and EGR1) were predicted at CpG island2.The results indicated that the differential expression of UCP3 gene in subcutaneous adipose tissue of Bama and Tibetan pigs might be caused by the different methylation levels of CpG island2.The higher DNA methylation of adipose tissue in Tibetan pigs might inhibit the expression of UCP3 gene at mRNA level by inhibiting the binding of transcription factors to promoter regulatory regions.

Key words: Bama pigs; Tibetan pigs; UCP3 gene; adipose tissue; methylation

CLC Number:

FAN Yiping, WANG Yanfang, TAO Cong. Expression and Methylation Analysis of UCP3 Gene in Adipose Tissue of Bama and Tibetan Pigs[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(12): 3627-3634.

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Expression and Methylation Analysis of UCP3 Gene in Adipose Tissue of Bama and Tibetan Pigs

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