KDM2B基因克隆及其在牦牛组织和卵母细胞减数分裂过程中的表达研究

doi:10.11843/j.issn.0366-6964.2018.03.009

Abstract

Abstract:

This research was conducted to clone the Lysine-specific histone demethylase 2B (KDM2B) gene in yak, identify its expression pattern in various tissues and meiosis process of oocytes, respectively, which might provide reliable basis for studying the mechanism of KDM2B in oocyte meiosis of yak. The samples of yak heart, liver, spleen, lung, kidney, brain, small intestine, stomach, muscle, ovary, testicle and uterus were collected after slaughtering. Total RNAs in different samples were extracted.Ovaries from healthy yaks of 3 to 5 years old were selected for collecting the cumulus-oocyte complex (COCs), and then oocytes and cumulus cells were collected by hyaluronidase. The coding sequence of KDM2B gene was cloned by RT-PCR. The mRNA expression of KDM2B in different tissues was determined by quantitative real-time PCR (qRT-PCR). The COCs in 3 stages of GV, MI and MⅡ were cultured in vitro, and the expression of KDM2B in the process of oocyte meiosis was detected by qRT-PCR. The results showed that the CDS region of KDM2B was 3 930 bp, encoding 1 309 amino acids. Compared with the existing predicted yak sequence, it belonged to long-form transcript and had high homology with cattle, sheep and goat, but had lower homology with zabra, fish and chicken. KDM2B gene of yak was widely expressed in various tissues, and its expression was relatively higher in spleen, uterus, testicle and ovary. The expression level of KDM2B in MI stage was significantly higher than that in GV and MⅡ stage in oocytes (P<0.01). In cumulus granulosa cells, the expression level of KDM2B was significantly increased by the process of oocyte meiosis (P<0.01). This research provided basic data for further research of oocyte meiosis mechanism and improving breeding efficiency in yak.

Key words: yak, Lysine-specific histone demethylase 2B gene, tissue expression, oocyte, meiosis

CLC Number:

S823.8⁺5.2

CAI Wen-yi, XIONG Xian-rong, CHEN Tong, YANG Xian-ying, HAN Jie, AGUO Yue-da, LI Jian. Molecular Cloning of KDM2B Gene and Its Expression Pattern in Tissues and the Process of Oocyte Meiosis in Yak[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(3): 534-541.

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Molecular Cloning of KDM2B Gene and Its Expression Pattern in Tissues and the Process of Oocyte Meiosis in Yak

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