牦牛HDAC1基因克隆及其在组织和卵母细胞减数分裂过程中的表达研究

doi:10.11843/j.issn.0366-6964.2019.10.005

畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (10): 1997-2004.doi: 10.11843/j.issn.0366-6964.2019.10.005

牦牛HDAC1基因克隆及其在组织和卵母细胞减数分裂过程中的表达研究

王斌, 殷实, 熊显荣, 秦文昌, 黄向月, 李键*

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, 成都 610041

收稿日期:2019-03-11 出版日期:2019-10-23 发布日期:2019-10-23
通讯作者: 李键,主要从事牦牛细胞生物学和发育生物学研究,E-mail:jianli_1967@163.com
作者简介:王斌(1993-),男,山西原平人,硕士生,主要从事动物细胞与胚胎工程研究,E-mail:15708451401@163.com
基金资助:
中央高校基本科研业务费专项基金项目（2018NQN33）；四川省科技支撑计划（2017NZ0076）；牦牛遗传资源与保护创新团队（13CXTD01）；西南民族大学研究生创新型科研项目（CX2018SZ10）；青藏高原动物遗传资源保护与利用重点实验室资助项目

Cloning of Yak HDAC1 Gene and Its Expression Pattern in Tissues and Oocyte during the Process of Meiosis

WANG Bin, YIN Shi, XIONG Xianrong, QIN Wenchang, HUANG Xiangyue, LI Jian*

Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation of Ministry of Education, College of Life Science and Technology, Southwest Minzu University, Chengdu 610041, China

Received:2019-03-11 Online:2019-10-23 Published:2019-10-23

摘要/Abstract

摘要： 旨在克隆牦牛组蛋白去乙酰化酶1（histone deacetylases 1，HDAC1）基因，并检测其在牦牛不同组织及卵母细胞减数分裂过程中的表达水平，从而为研究HDAC1在牦牛生殖发育中的作用机制提供理论依据。本试验采用牦牛为研究对象，通过RT-PCR技术获得牦牛HDAC1基因CDS序列，使用相关生物信息学软件分析其结构和功能，通过实时荧光定量PCR（quantitative real-time PCR，qRT-PCR）检测HDAC1基因在牦牛脑、心、肌肉、卵巢、肾、脾、小肠、肝、肺和子宫组织中的表达谱及在卵母细胞减数分裂过程中mRNA的表达水平。结果表明，HDAC1基因开放阅读框为1 302 bp，编码433个氨基酸，与黄牛、山羊和绵羊的同源性较高；HDAC1基因在牦牛的各个组织中均有表达，其中在脾、肾和小肠中的表达量极显著高于其他组织（P<0.01）。在牦牛减数第一次分裂中期（MⅠ）和减数第二次分裂中期（MⅡ）卵母细胞中，HDAC1基因的表达水平极显著高于生发泡（germinal vesicle，GV）期（P<0.01）。提示，HDAC1基因参与牦牛卵母细胞减数分裂过程。本试验为进一步研究HDAC1基因在高寒、低氧环境下的牦牛生殖繁育中的作用提供了理论依据。

Abstract: The objective of this research was to clone the histone deacetylases 1 (HDAC1) gene of yak and identify its expression patterns in various tissues and oocytes during meiosis process, respectively, which would provide theoretical basis for studying the role mechanism of HDAC1 in reproductive development of yak. The samples of yak heart, liver, spleen, lung, kidney, small intestine, brain, muscle, ovary and uterus were collected after slaughtering. Total RNAs in different samples were extracted. The coding sequence of HDAC1 gene was cloned by RT-PCR. Its structure and function were analyzed by bioinformatics softwares. The mRNA expression of HDAC1 in different tissues and oocyte during meiosis process was detected by quantitative real-time PCR (qRT-PCR). The results showed that the open reading frame HDAC1 was 1 302 bp, encoding 433 amino acids. It had high homology with cattle, sheep and goat. HDAC1 gene of yak was widely expressed in various tissues, the expression level of which was significantly higher in spleen, kidney and small intestine(P<0.01). The expression level of HDAC1 in MI and MⅡ stages was significantly higher than that in GV stage in oocytes of yak(P<0.01). This study suggested that HDAC1 was involved in meiosis process of oocytes in yak, which provided basic data for studying the role of HDAC1 in reproductive development of yak under the harsh natural environment of the plateau.

中图分类号:

S823.85

王斌, 殷实, 熊显荣, 秦文昌, 黄向月, 李键. 牦牛HDAC1基因克隆及其在组织和卵母细胞减数分裂过程中的表达研究[J]. 畜牧兽医学报, 2019, 50(10): 1997-2004.

WANG Bin, YIN Shi, XIONG Xianrong, QIN Wenchang, HUANG Xiangyue, LI Jian. Cloning of Yak HDAC1 Gene and Its Expression Pattern in Tissues and Oocyte during the Process of Meiosis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(10): 1997-2004.

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牦牛HDAC1基因克隆及其在组织和卵母细胞减数分裂过程中的表达研究

Cloning of Yak HDAC1 Gene and Its Expression Pattern in Tissues and Oocyte during the Process of Meiosis

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