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

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

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、MⅠ和MⅡ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,MⅡ期表达水平最高,极显著高于GV和MⅠ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。

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, MⅠ and MⅡ 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 MⅡ stage was the highest, which was significantly higher than GV and MⅠ 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.