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2025年4月6日 星期日
农业生物技术学报  2020, Vol. 28 Issue (6): 984-993    DOI: 10.3969/j.issn.1674-7968.2020.06.004
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
天祝白牦牛ACACA基因组织表达及脂肪组织甲基化模式分析
石学红1, 石斌刚1, 左志1, 赵世杰1, 何兆华1, 张雪萍1, 赵志东1, 王继卿1, 周智德2, 胡江1,*
1 甘肃农业大学 动物科学技术学院/甘肃省草食动物生物技术重点实验室/甘肃省牛羊基因改良工程实验室,兰州 730070;
2 甘肃省天祝藏族自治县松山镇畜牧兽医站,天祝 733200
Tissue Expression of ACACA Gene and Methylation Pattern Analysis of Adipose Tissue in Tianzhu White Yak (Bos grunniens)
SHI Xue-Hong1, SHI Bin-Gang1, ZUO Zhi1, ZHAO Shi-Jie1, HE Zhao-Hua1, ZHANG Xue-Ping1, ZHAO Zhi-Dong1, WANG Ji-Qing1, ZHOU Zhi-De2, HU Jiang1,*
1 Faculty of Animal Science and Technology, Gansu Agricultural University/Gansu Key Laboratory of Herbivorous Animal Biotechnology/Gansu Engineering Lab of Genetic Improvement in Ruminants, Lanzhou 730070, China;
2 Tianzhu County Songshan Town Animal Husbandry Veterinary Station, Tianzhu 733200, China
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摘要 乙酰辅酶A羧化酶α (acetyl-CoA carboxylases alpha, ACACA)是脂肪酸从头合成的关键限速酶,在脂肪酸生物合成中起重要调控作用。本研究以0.5和4.5岁天祝白牦牛(Bos grunniens)为研究对象,采用实时荧光定量PCR (quantitative real-time PCR, qRT-PCR)和重亚硫酸盐测序(bisulfite sequencing PCR, BSP)方法,检测牦牛ACACA基因在心脏、肝脏、皮下脂肪等13个组织中的表达量,以及P启动子区CpG岛在皮下脂肪组织中的甲基化水平,研究该基因组织表达规律及皮下脂肪组织中的表观遗传机制。结果表明,ACACA基因在牦牛上的表达量存在组织和年龄差异,0.5岁牦牛肝脏、脑、背最长肌和肾脏中高度表达且肝脏中表达量显著高于其他组织(P<0.05),4.5岁牦牛皮下脂肪和肝脏中表达量极显著高于其他组织(P<0.01);0.5岁牦牛肾脏中表达量极显著高于4.5岁牦牛(P<0.01),而在皮下脂肪、肝脏和睾丸中表达量显著或极显著低于4.5岁牦牛(P<0.05或P<0.01)。皮下脂肪组织中,ACACA基因P启动子第1 CpG岛区(g.-795 bp~-193 bp)高度去甲基化且存在年龄差异,且BSP-1引物扩增区平均甲基化水平0.5岁牦牛极显著高于4.5岁牦牛(P<0.01);0.5岁牦牛BSP-1引物扩增区CpG11、CpG31、CpG35、CpG37位点及BSP-2引物扩增区CpG5'、CpG13'、CpG15'、CpG27'、CpG28'位点甲基化水平显著或极显著高于4.5岁牦牛(P<0.05或P<0.01);P启动子第1 CpG岛区甲基化水平与该基因在皮下脂肪组织中的表达量呈负相关。本研究揭示了不同年龄段牦牛ACACA基因组织表达和P启动子甲基化差异,为牦牛分子遗传研究提供理论基础。
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石学红
石斌刚
左志
赵世杰
何兆华
张雪萍
赵志东
王继卿
周智德
胡江
关键词 乙酰辅酶A羧化酶α基因组织表达皮下脂肪组织甲基化天祝白牦牛    
Abstract:Acetyl-CoA carboxylases alpha (ACACA) is a key rate-limiting enzyme in the de novo synthesis of fatty acids, which plays an important role in the regulation of fatty acid biosynthesis. In this study, 0.5-year-old and 4.5-year-old Tianzhu white yaks (Bos grunniens) were studied using quantitative real-time PCR (qRT-PCR) and heavy sulfite sequencing (BSP) methods. The expression of ACACA gene in 13 tissues such as heart, liver, and subcutaneous fat, as well as the methylation level of CpG island in P promoter region in subcutaneous adipose tissue were detected, and the gene expression regular and epigenetic mechanism in subcutaneous adipose tissue were studied. The results showed that the expression of ACACA gene in yaks were different in tissues and ages.The liver, brain, dorsal longest muscle and kidney tissues of 0.5-year-old yaks were highly expressed and the liver expression was significantly higher than other tissues (P<0.05);The expression of subcutaneous adipose and liver in 4.5-year-old yaks was extremely significantly higher than other tissues (P<0.01); The expression in the kidney of 0.5-year-old yaks were extremely significantly higher than that of 4.5-year-old yaks (P<0.01), but the expression of subcutaneous adipose, liver and testicular was significantly or extremely significantly less than that 4.5-year-old yaks (P<0.05 or P<0.01).In the subcutaneous adipose tissue, the ACACA gene P promoter 1st CpG island area (g.-795 bp~-193 bp) was highly demethylated and there were age differences. The average methylation level of BSP-1 primer amplification zone in 0.5-year-old yaks was extremely significantly higher than that in 4.5-year-old yaks (P<0.01). 0.5-year-old yaks BSP-1 primer amplification zone CpG11, CpG31, CpG35, CpG37 site and BSP-2 primer amplification zone CpG5', CpG13', CpG15', CpG27', CpG28' site methylation level significantly or extremely significantly higher than 4.5-year-old yaks (P<0.05 or P< 0.01); The level of methylation in the 1st CpG island region of the P promoter was negatively correlated with the expression of the gene in subcutaneous adipose tissue. This study revealed the differences of ACACA gene expression and P promoter methylation in yaks of different ages, which provide theoretical basis for molecular genetic research of yaks.
Key wordsACACA gene    Tissue expression    Subcutaneous fatty tissue    Methylation    Tianzhu white yak
收稿日期: 2019-10-21     
ZTFLH:  S829.9  
基金资助:甘肃省基础研究创新群体项目(17JR5RA137); 甘肃省农业生物技术研究与应用开发项目(GNSW-2016-29); 国家自然科学基金(31160451)
通讯作者: *,huj@gsau.edu.cn   
引用本文:   
石学红, 石斌刚, 左志, 赵世杰, 何兆华, 张雪萍, 赵志东, 王继卿, 周智德, 胡江. 天祝白牦牛ACACA基因组织表达及脂肪组织甲基化模式分析[J]. 农业生物技术学报, 2020, 28(6): 984-993.
SHI Xue-Hong, SHI Bin-Gang, ZUO Zhi, ZHAO Shi-Jie, HE Zhao-Hua, ZHANG Xue-Ping, ZHAO Zhi-Dong, WANG Ji-Qing, ZHOU Zhi-De, HU Jiang. Tissue Expression of ACACA Gene and Methylation Pattern Analysis of Adipose Tissue in Tianzhu White Yak (Bos grunniens). 农业生物技术学报, 2020, 28(6): 984-993.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2020.06.004     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2020/V28/I6/984
 
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