华北农学报 ›› 2020, Vol. 35 ›› Issue (2): 210-216. doi: 10.7668/hbnxb.20190532

所属专题: 畜牧 生物技术 热点文章

• 畜牧·兽医 • 上一篇    下一篇

牦牛HDAC8基因特征及其在组织和卵母细胞成熟过程中的表达分析

秦文昌, 殷实, 李泽沛, 王斌, 杨柳青, 周婧雯, 李键   

  1. 西南民族大学 生命科学与技术学院, 青藏高原动物遗传资源保护与利用教育部重点实验室, 青藏高原动物遗传资源保护与利用四川省重点实验室, 四川 成都 610041
  • 收稿日期:2019-11-07 出版日期:2020-04-28
  • 通讯作者: 李键(1967-),男,四川理县人,教授,博士,硕士生导师,主要从事牦牛细胞生物学和发育生物学研究;殷实(1988-),男,河南焦作人,讲师,博士,主要从事动物细胞与胚胎工程研究。
  • 作者简介:秦文昌(1994-),男,青海民和人,硕士,主要从事动物细胞与胚胎工程研究。
  • 基金资助:
    国家重点研发专项(2018YFD0502304);四川省科技支撑计划(2017NZ0076);牦牛遗传资源与保护创新团队(13CXTD01);青藏高原生态畜牧业协同创新中心开放基金(QZGYXT05);中央高校基本科研业务费专项基金(2018NQN33)

The Characteristic of Yak HDAC8 Gene and Analysis of Their Expression in Different Tissues and During the Oocyte Maturation

QIN Wenchang, YIN Shi, LI Zepei, WANG Bin, YANG Liuqing, ZHOU Jingwen, LI Jian   

  1. College of Life Science and Technology, Southwest Minzu University, Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Key Laboratory of Sichuan Province for Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation, Chengdu 610041, China
  • Received:2019-11-07 Published:2020-04-28

摘要: 为探讨组蛋白去乙酰化酶8(Histone deacetylases 8,HDAC8)在牦牛生殖中的作用,以牦牛肝脏cDNA为模板,采用RT-PCR方法克隆牦牛HDAC8基因,并通过生物信息学方法分析牦牛HDAC8序列;利用实时荧光定量PCR(Quantitative Real-time PCR,qRT-PCR)技术检测该基因在牦牛不同组织及卵母细胞体外成熟过程(GV、M Ⅰ和M Ⅱ期)中的表达规律。结果表明,牦牛HDAC8基因序列为1 109 bp(GenBank No.MK889494),CDS为1 008 bp,编码氨基酸335个;序列比对分析发现,牦牛与其他哺乳动物的相似性在94%以上,表明HDAC8在哺乳类动物进化过程及蛋白结构水平上具有较高的保守性;组织表达分析发现,HDAC8在肝脏组织中的表达水平最高,极显著高于其他组织(P<0.01);HDAC8基因在卵母细胞不同发育阶段中具有不同的表达模式,表达水平随着卵母细胞成熟阶段的递增呈上升趋势,M Ⅱ期表达水平最高,极显著高于GV和M Ⅰ期(P<0.01)。综上,HDAC8基因可能在牦牛肝脏代谢及卵母细胞的成熟过程中起一定的调控作用,为进一步探索HDAC8在牦牛生殖中的调控作用提供一定的理论基础。

关键词: 牦牛, HDAC8基因, 组织表达, 卵母细胞

Abstract: In order to explore the role of HDAC8 in the reproductive process of yak,the yak HDAC8 gene sequence was firstly cloned using cDNA as a template from liver by RT-PCR and then was analyzed by bioinformatics; the expression of HDAC8 gene in different tissues and in the process during oocyte in vitro maturation (GV, M Ⅰ and M Ⅱ phases) was detected by quantitative Real-time PCR (qRT-PCR). The results showed that the HDAC8 gene sequence was 1 109 bp (GenBank No.MK889494), CDS was 1 008 bp, and 335 amino acids were encoded. Sequence alignment analysis showed that the similarity between yak and other mammals was over 94%, indicating that HDAC8 was highly conserved in mammalian evolution and protein structure. The tissues expression analysis showed that HDAC8 highly expressed in liver, which was remarkably higher than other tissues (P<0.01). The expression pattern of HDAC8 gene was different in oocyte of yak in different stages. The expression level of HDAC8 mRNA in oocyte increased with the increasing stage of oocyte maturation, and the expression level of M Ⅱ stage was the highest, which was significantly higher than GV and M Ⅰ stages (P<0.01). These data indicated that HDAC8 gene might play a regulatory role in the liver metabolism and during the oocyte maturation in yak. This study providing a theoretical basis for further exploration of the regulation of HDAC8 in reproductive of yak.

Key words: Yak, HDAC8, Tissue expression, Oocyte

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引用本文

秦文昌, 殷实, 李泽沛, 王斌, 杨柳青, 周婧雯, 李键. 牦牛HDAC8基因特征及其在组织和卵母细胞成熟过程中的表达分析[J]. 华北农学报, 2020, 35(2): 210-216. doi: 10.7668/hbnxb.20190532.

QIN Wenchang, YIN Shi, LI Zepei, WANG Bin, YANG Liuqing, ZHOU Jingwen, LI Jian. The Characteristic of Yak HDAC8 Gene and Analysis of Their Expression in Different Tissues and During the Oocyte Maturation[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(2): 210-216. doi: 10.7668/hbnxb.20190532.

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