Analysis of the Bovine DLK1 Gene Polymorphism and Its Relation to Lipid Metabolism in Chinese Simmental
Analysis of the Bovine DLK1 Gene Polymorphism and Its Relation to Lipid Metabolism in Chinese Simmental
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中国西门塔尔牛DLK1基因多态性分析及其与脂质代谢的关系
DOI：
10.3390/ani10060923
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发表时间：
2020-05
期刊：
影响因子：
--
通讯作者：
Zhihui Zhao
中科院分区：
文献类型：
--
作者：
Mengyan Wang;Ping Jiang;Xiang Yu;Jiaqi Mi;Zitong Bai;Xiuqi Zhang;Yinuo Liu;Xibi Fang;Runjun Yang;Zhihui Zhao
Delta-like non-canonical Notch ligand 1 （DLK1）是一个与脂质代谢相关的候选基因。为了验证DLK1基因在中国西门塔尔牛脂质代谢中的作用，我们检测了DLK1基因对牛胎成纤维细胞（BFFs）脂质代谢的影响。同时，我们还检测了牛DLK1单核苷酸多态性位点（SNPs）与中国西门塔尔牛胴体脂肪覆盖率、腰眼肌面积、脂肪颜色评分等经济性状和脂肪酸组成的关系。在本研究中，我们精确构建并转染了DLK1基因在闺蜜中的过表达和干扰载体。结果表明，过表达DLK1基因可降低细胞内甘油三酯（TGs）含量，而干扰DLK1基因可增加细胞内甘油三酯（TGs）含量。脂肪酸组成的气相色谱分析表明，随着DLK1表达的改变，辛酸酸和γ-亚麻酸的含量受到调节。采用限制性内切片段长度多态性（RPLF-PCR）检测方法，在中国西门塔尔植物中发现了两个snp和与这些性状相关的基因。综上所述，我们从细胞水平到群体遗传多态性验证了牛DLK1对脂肪酸代谢的影响，这也为进一步研究DLK1基因对体内脂质代谢的影响铺平了道路。摘要本研究通过在bff中精确构建和转染DLK1基因过表达和干扰载体，研究DLK1基因在体外脂质代谢中的作用。与对照组相比，pGPU6-shDLK1组DLK1 mRNA和蛋白水平表达有降低的趋势，TGs显著升高（p < 0.05）。与对照组相比，pBI-CMV3-DLK1组DLK1 mRNA和蛋白表达水平均升高，TGs含量显著降低（p < 0.05）。同时，我们采用限制性内切片段长度多态性（RFLP-PCR）检测方法进一步筛选snp，探索分析该基因与28月龄中国西门塔尔经济性状及牛最长肌脂肪酸组成的关系。结果表明，IVS3 + 478 C > T和IVS3 + 609 T >g两个snp与中国西门犀胴体和肉质性状（胴体脂肪覆盖率、腰眼肌面积和脂肪色评分）显著相关。综上所述，DLK1可影响牛脂质代谢，这两个snp可作为肉牛肉质性状的遗传标记。
Simple Summary Delta-like non-canonical Notch ligand 1 (DLK1) is a candidate gene associated with lipid metabolism. In order to verify the function of the DLK1 gene on lipid metabolism in Chinese Simmental cattle, we identified the effect of DLK1 on lipid metabolism in bovine fetal fibroblast cells (BFFs). At the same time, we also detected the relationship between single-nucleotide polymorphism sites (SNPs) of bovine DLK1 with the economic traits and fatty acids composition in Chinese Simmental cattle, such as the carcass fat coverage rate, loin eye muscle area, and fat color score, etc. In the present study, we precisely constructed and transfected the overexpression and interference vectors of the DLK1 gene in BFFs. the results showed that the overexpression of the DLK1 gene could decrease the contents of intracellular triglycerides (TGs) while interference of the DLK1 gene could increase the TGs contents. Gas chromatography analysis of the fatty acid composition showed that the contents of octanoate acid and γ-linolenate acid were regulated as the expression of DLK1 was altered. We found two SNPs and genes associated with these traits in Chinese Simmental by the restriction fragment length polymorphism (RPLF-PCR) detection method. In summary, we verified the effects of bovine DLK1 on fatty acid metabolism from the cellular level to population genetic polymorphism, which also paved the way for further studying of the effects of the DLK1 gene on lipid metabolism in vivo. Abstract In this study, we precisely constructed and transfected the overexpression and interference vectors in BFFs to evaluate the role of DLK1 gene on lipid metabolism in vitro. The expression of of DLK1 in the mRNA and protein level tended to reduce, and TGs were significantly increased in the pGPU6-shDLK1 group compared to the control group (p < 0.05). The expression of DLK1 in the mRNA and protein level were increased in the pBI-CMV3-DLK1 group compared to the control group, and the TGs content showed a significant decrease in the pBI-CMV3-DLK1 group (p < 0.05). Meanwhile, we used the restriction fragment length polymorphism (RFLP-PCR) detection method to screen SNPs further to explore and analyze the relationship between the gene and the economic traits of 28-month-old Chinese Simmental and the fatty acids composition of cattle longissimus muscle. The result showed that two SNPs, IVS3 + 478 C > T and IVS3 + 609 T > G, were identified as being significantly associated with carcass and meat quality traits in Chinese Simmental, such as the carcass fat coverage rate, loin eye muscle area, and fat color score. In summary, our results indicated that DLK1 can affect lipid metabolism in bovine and these two SNPs might be applied as genetic markers of meat quality traits for beef cattle breeding.
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DOI：
10.1016/s1096-4959(02)00265-8
发表时间：
2003-02
期刊：
Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology
影响因子：
--
作者：
T. Vuocolo;R. Pearson;P. Campbell;R. Tellam
通讯作者：
T. Vuocolo;R. Pearson;P. Campbell;R. Tellam
影响因子：
1.9
作者：
S. Phinney
通讯作者：
S. Phinney
影响因子：
2.5
作者：
Kim, KS;Kim, LL;Rothschild, MF
通讯作者：
Rothschild, MF
DOI：
10.1073/pnas.1406119111
发表时间：
2014-11-11
影响因子：
11.1
作者：
Charalambous, Marika;Da Rocha, Simao Teixeira;Ferguson-Smith, Anne C.
通讯作者：
Ferguson-Smith, Anne C.
影响因子：
9.2
作者：
Davis, E;Jensen, CH;Charlier, C
通讯作者：
Charlier, C