牦牛TSHB、DIO2和DIO3基因的克隆及其在繁殖轴的表达研究

doi:10.13560/j.cnki.biotech.bull.1985.2018-0793

摘要/Abstract

摘要： 为了解TSHB-DIO2/DIO3系统对牦牛季节性发情的调控机制,以牦牛为研究对象,运用RT-PCR方法克隆牦牛TSHB、DIO2、DIO3基因的cDNA序列,结合GenBank已公布的普通牛TSHB、DIO2、DIO3基因序列,构建遗传进化树,采用荧光定量 PCR 技术检测其在牦牛下丘脑、垂体、卵巢、输卵管、子宫中的表达水平。结果表明,牦牛TSHB、DIO2、DIO3基因均与普通牛有最近的分子进化关系;TSHB基因CDS区为417 bp,在垂体中的表达水平高于其他组织,差异极显著（P<0.01）;DIO2基因cDNA 为951 bp,在所检测的组织中均有表达,且在子宫中的表达水平高于其他组织,差异极显著（P<0.01）;DIO3基因cDNA为873 bp,在子宫组织中未检测到表达,在卵巢中的表达水平高于垂体、下丘脑和输卵管,差异极显著（P<0.01）。提示TSHB-DIO2/DIO3可能参与牦牛繁殖调控,旨为揭示牦牛季节性繁殖分子调控机制提供参考。

关键词: TSHB, DIO2, DIO3, 牦牛, 组织表达

Abstract: In order to study the regulation mechanism of TSHB-DIO2/DIO3 system on seasonal breeding of the yak,RT-PCR was used to analyze the cDNA sequences of gene TSHB,DIO2,and DIO3,and the phylogenetic tree was constructed based on the sequences of bovine gene TSHB,DIO2,and DIO3 in the GenBank. Further,their mRNA expression levels in hypothalamus,pituitary,ovary,oviduct and uterus were analyzed by real-time PCR. These results indicated that there was very close evolutionary relationship in yak’s gene TSHB,DIO2,and DIO3 between a yak and a bovine. The coding region of gene TSHB was 417 bp,and its expression level in pituitary was significantly higher than that in other tissues（P < 0.01）. The cDNA sequence of DIO2 gene was 951 bp and expressed in all tissues,but it was significantly higher in the uterus than that in the other four tissues（P < 0.01）. The cDNA sequences of DIO3 gene was 873 bp and was not detected in the uterus,the expression of which in the ovary was significantly higher than that in the pituitary,hypothalamus and oviduct（P < 0.01）. This study suggests that the TSHB-DIO2/DIO3 may be involved in the regulation of yak reproduction,which may provide a reference for preliminarily revealing the molecular regulation mechanism of yak seasonal breeding.

Key words: TSHB, DIO2, DIO3, yak, tissue expression

王琴, 卢建远, 字向东. 牦牛TSHB、DIO2和DIO3基因的克隆及其在繁殖轴的表达研究[J]. 生物技术通报, 2019, 35(3): 132-137.

WANG Qin, LU Jian-yuan, ZI Xiang-dong. Cloning of Gene TSHB,DIO2 and DIO3 and Their Expression Levels in Reproductive Axis of Female Yaks[J]. Biotechnology Bulletin, 2019, 35(3): 132-137.

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