畜牧兽医学报2018，Vol.49Issue(3)：534-541,8.DOI:10.11843/j.issn.0366-6964.2018.03.009
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
Molecular Cloning of KDM2B Gene and Its Expression Pattern in Tissues and the Process of Oocyte Meiosis in Yak
摘要
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.关键词
牦牛/KDM2B基因/组织表达/卵母细胞/减数分裂Key words
yak/Lysine-specific histone demethylase 2B gene/tissue expression/oocyte/meiosis分类
农业科技引用本文复制引用
蔡雯祎,熊显荣,陈通,杨显英,韩杰,阿果约达,李键..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[J].畜牧兽医学报,2018,49(3):534-541,8.基金项目
四川省科技支撑计划(2017NZ0076) （2017NZ0076）
西南民族大学创新团队(2015NZYTD02) （2015NZYTD02）
四川省肉牛创新团队资助 （）
