Cloning on CDS of Yak (Bos grunniens) KDM1B Gene and Its Expression in Different Tissues and During the Oocyte Maturation
QIN Wen-Chang, YIN Shi, XIONG Xian-Rong, WANG Bin, UANG Xiang-Yue, ZHOU Jing-Wen,LI Jian
1 College of Life Science and Technology, Southwest Minzu University, Chengdu 610041, China; 2 Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, Chengdu 610041, China
Abstract：Lysine-specific histone demethylase 1B (KDM1B) is required for de novo methylation of some imprinted genes in oocytes. The aim of this study was to clone the CDS sequence of the KDM1B gene of yak (Bos grunniens), and to analyze the expression characteristics of KDM1B in various tissues and the process of in vitro maturation of oocytes in yak. In this study, the CDS region of KDM1B was amplified from yak ovary cDNA by PCR.The physic-chemical properties, structure and conservation of KDM1B were predicted by bioinformatics. The expression of KDM1B in different tissues and in the process during oocyte in vitro maturation was detected by qRT-PCR. Result showed that the CDS of yak KDM1B gene was 1 737 bp (GenBank No. MK806390), encoding 578 amino acids. The molecular weight of protein encoded by KDM1B gene in yak was 64.90 kD. The theory isoelectric point was 7.20, and the grand average of hydroparthicity was -0.190, and the instability index was 41.73, predicted that KDM1B was a hydrophilic protein; The secondary structure of KDM1B consisted of α-helix, β-corner, and random coils. The amino acids sequence of KDM1B contains N-terminal double zinc finger (zf-CW), N-terminal SWIRM (Swi3p/Rsc8p/Moira, SWIRM) and C-terminal amine oxidase (AO) domain. The protein was predicted to be lacking signal peptides and cross membrane. Moreover, the conservation analysis showed that yak KDM1B gene had high homology comparison of mammals such as zebu (B. indicus) and cattle (B. taurus). The expression of KDM1B mRNA was found in various tissues of yak, and the relative expression was highest in small intestine. During in vitro maturation (GⅤ phase, MⅠphase, MⅡphase) of oocytes, the mRNA expression level of KDM1B was extremely significantly higher in MⅡ oocytes than that in GⅤ and MⅠ stages (P＜0.01), but no significant difference between GⅤ and MⅠ stages. These results would be beneficial for studying the role of KDM1B in the reproductive process in yak.
秦文昌, 殷实, 熊显荣, 王斌, 黄向月, 周婧雯, 李键. 牦牛KDM1B基因CDS区克隆及其在不同组织和卵母细胞成熟过程中的表达[J]. 农业生物技术学报, 2019, 27(9): 1604-1613.
QIN Wen-Chang, YIN Shi, XIONG Xian-Rong, WANG Bin, UANG Xiang-Yue, ZHOU Jing-Wen,LI Jian. Cloning on CDS of Yak (Bos grunniens) KDM1B Gene and Its Expression in Different Tissues and During the Oocyte Maturation. 农业生物技术学报, 2019, 27(9): 1604-1613.
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