华北农学报 ›› 2025, Vol. 40 ›› Issue (3): 199-206. doi: 10.7668/hbnxb.20195461

所属专题: 畜牧

• 畜牧·水产·兽医 • 上一篇    下一篇

奶山羊SIRT7基因序列特征、组织表达及其对乳脂合成的影响

李忠莹1,2, 郭震楠1, 朱洪林1, 李佳琳1, 施会彬1, 韩浩园1, 李君1, , 权凯1,2,   

  1. 1 河南牧业经济学院 动物科技学院,河南 郑州 450046
    2 宁夏大学 动物科技学院,宁夏 银川 750021
  • 收稿日期:2024-09-27 出版日期:2025-06-28
  • 通讯作者:
    李 君(1985—),女,河南虞城人,副教授,博士,主要从事动物遗传育种与泌乳调控研究。
    权 凯(1977—),男,甘肃陇西人,教授,博士,主要从事羊遗传育种与繁殖研究。
  • 作者简介:

    李忠莹(1996—),女,四川泸县人,在读硕士,主要从事动物遗传育种研究。

  • 基金资助:
    河南省自然科学基金项目(242300420148); 河南省重点研发与推广专项(科技攻关)(232102110007); 河南省现代农业产业技术体系建设专项资金项目(HARS-22-15-S)

Sequence Characteristics,Tissue Expression and Effect on Milk Fat Synthesis of SIRT7 Gene in Dairy Goats

LI Zhongying1,2, GUO Zhennan1, ZHU Honglin1, LI Jialin1, SHI Huibin1, HAN Haoyuan1, LI Jun1, , QUAN Kai1,2,   

  1. 1 College of Animal Science and Technology,Henan University of Animal Husbandry and Economy,Zhengzhou 450046,China
    2 School of Animal Science and Technology,Ningxia University,Yinchuan 750021,China
  • Received:2024-09-27 Published:2025-06-28

摘要:

旨在克隆奶山羊沉默信息调节因子2相关酶7基因(SIRT7)并进行生物信息学分析,检测该基因在奶山羊不同组织中的表达规律及其对奶山羊乳腺上皮细胞乳脂合成的影响。结果表明,通过设计SIRT7基因引物,成功克隆奶山羊SIRT7基因序列1 376 bp,编码区长1 203 bp,编码400个氨基酸,与绵羊同源性最高。SIRT7是亲水性蛋白,不含信号肽和跨膜结构,二级结构有无规则卷曲、α-螺旋、延伸链和β-转角。qRT-PCR结果显示,SIRT7基因在奶山羊肝脏组织中的相对表达量最高,在其他组织中也有一定的表达量。构建SIRT7基因过表达载体pcDNA3.1-SIRT7,将其转染至奶山羊乳腺上皮细胞中,与空载体对照组相比,过表达组SIRT7基因mRNA水平上调约33倍。SIRT7基因过表达后,乳脂代谢相关基因中硬脂酰辅酶A去饱和酶基因(SCD1)表达量显著降低,抗原分化簇36基因(CD36)、二酰基甘油酰基转移酶1基因(DGAT1)、固醇调节元件结合蛋白1基因(SREBP1)的表达量极显著降低;细胞中甘油三酯含量显著降低;细胞核周脂滴聚集量明显减少。因此,SIRT7基因抑制奶山羊乳腺上皮细胞的脂质代谢,为进一步探索SIRT7在奶山羊乳腺上皮细胞中的调控机制奠定基础。

关键词: 奶山羊, SIRT7, 基因克隆, 组织表达, 乳脂合成

Abstract:

The aim of this study was to clone the silent mating type information regulator 2-related enzyme 7(SIRT7)gene,detect its expression pattern in different tissues and investigate its effect on milk fat synthesis in mammary epithelial cells of dairy goats.The results showed that by designing the primers of SIRT7 gene,the sequence of SIRT7 gene in dairy goat was successfully cloned,with 1 376 bp and a coding region of 1 203 bp,encoding 400 amino acids and showing the highest homology with sheep.SIRT7 was a hydrophilic protein,free of signal peptides and transmembrane structures,with secondary structures such as irregular coils,α-helices,extended chains and β-turns.The relative expression of SIRT7 gene was the highest in the liver tissues of dairy goats,and it also had certain expression levels in other tissues by qRT-PCR.The pcDNA3.1-SIRT7 overexpression vector was successfully constructed and transfected into mammary epithelial cells of dairy goats.Compared with the control group,the mRNA level of SIRT7 gene in the overexpression group was up-regulated about 33 times.After SIRT7 gene overexpression,the expression of stearoyl-CoA desaturase-1(SCD1)gene was significantly decreased,and the expression levels of cluster of differentiation 36(CD36),diacylglycerol acyltransferase 1(DGAT1),sterol regulatory element binding protein 1(SREBP1)genes extremely significantly reduced.The content of triglyceride in cells was significantly decreased.The results of oil red O staining showed that the accumulation of lipid droplets around the nucleus was significantly reduced.Thus, SIRT7 gene inhibited lipid metabolism in mammary epithelial cells of dairy goats,which laid a foundation for further exploration of the regulatory mechanism of SIRT7 in mammary epithelial cells of dairy goats.

Key words: Dairy goat, SIRT7, Gene cloning, Tissue expression, Milk fat synthesis

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引用本文

李忠莹, 郭震楠, 朱洪林, 李佳琳, 施会彬, 韩浩园, 李君, 权凯. 奶山羊SIRT7基因序列特征、组织表达及其对乳脂合成的影响[J]. 华北农学报, 2025, 40(3): 199-206. doi: 10.7668/hbnxb.20195461.

LI Zhongying, GUO Zhennan, ZHU Honglin, LI Jialin, SHI Huibin, HAN Haoyuan, LI Jun, QUAN Kai. Sequence Characteristics,Tissue Expression and Effect on Milk Fat Synthesis of SIRT7 Gene in Dairy Goats[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(3): 199-206. doi: 10.7668/hbnxb.20195461.