Identification and Analysis of a Functional SNP 1196C > A in 3 ' UTR of Chicken IGFBP2 Gene

文献类型： 外文期刊

第一作者： Yu Ying-Ying

作者： Yu Ying-Ying;Qiao Shu-Pei;Sun Ying-Ning;Song He;Zhang Xiao-Fei;Yan Xiao-Hong;Li Hui;Wang Ning;Yu Ying-Ying;Qiao Shu-Pei;Sun Ying-Ning;Song He;Zhang Xiao-Fei;Yan Xiao-Hong;Li Hui;Wang Ning;Yu Ying-Ying;Qiao Shu-Pei;Sun Ying-Ning;Song He;Zhang Xiao-Fei;Yan Xiao-Hong;Li Hui;Wang Ning;Sun Ying-Ning

作者机构：

关键词： chicken;IGFBP2 gene;SNP;3 ' UTR;gga-miR-456-3p

期刊名称：PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS （ 影响因子：0.351； 五年影响因子：0.272 ）

ISSN： 1000-3282

年卷期： 2014 年 41 卷 11 期

页码：

收录情况： SCI

摘要： We previously mapped a QTL significantly influencing the abdominal fat weight and the percentage of abdominal fat of chicken on chicken chromosome 7, and IGFBP2 is the only known gene located within this QTL region. We further found that the 1196C>A, a single nucleotide polymorphism (SNP) in 3'UTR of Chicken IGFBP2 gene, is significantly associated with chicken abdominal fat weight and percentage of abdominal fat. Bioinformatics analysis suggests that this SNP is located at a potential binding site of gga-miR-456-3p. We hypothesized that IGFBP2 may be a target gene of gga-miR-456-3p, and this SNP may affect the gga-miR-456-3p-mediated downregulation of IGFBP2. To test and decipher this hypothesis, in the present study we constructed the 3' UTR reporter vectors for the two individual alleles of the IGFBP2 SNP (1196C>A), and compared the reporter activity between the two individual alleles in both DF1 cells and chicken preadipocytes. Using the miRNA mimics and inhibitor of gga-miR-456-3p, we further assessed the effects of gga-miR-456-3p on the reporter activity of the two individual alleles of the IGFBP2 SNP and the endogenous IGFBP2 expression at mRNA and protein levels in DF1 cells. The reporter assay showed that in both DF1 cells and chicken preadipocytes, allele A had higher reporter activities at protein and mRNA levels than allele C, indicating that this SNP is a functional SNP. Further studies demonstrated that in DF1 cells, gga-miR-456-3p mimcs and inhibitor had effect on allele C reporter activity, but not on allele A reporter activity. Quantitative real-time RT-PCR and Western blot analyses showed that gga-miR-456-3p mimcs and inhibitor regulated the endogenous expression of chicken IGFBP2 gene at mRNA and protein levels. Taken together, our results demonstrated that IGFBP2 gene is a target gene of gga-miR-456-3p, and the SNP 1196C> A is a functional SNP. Our findings are of great significance to the marker-assisted selection for lean chicken and clarification of gene function and regulation of chicken IGFBP2 gene in chicken adipose development.

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