牦牛Bcl-2、Bax基因克隆及其在生殖轴上的表达分析

doi:10.16431/j.cnki.1671-7236.2019.07.001

摘要/Abstract

摘要：

试验旨在分析Bcl-2与Bax基因的序列特性，并分析其在母牦牛生殖轴上的表达特点，为探讨其在牦牛繁殖活动中的调控作用奠定基础。试验采集健康母牦牛与母黄牛下丘脑、垂体、卵巢、输卵管及子宫组织样品，通过RT-PCR扩增并克隆Bcl-2与Bax基因，并采用生物信息学软件进行序列分析；利用实时荧光定量PCR法检测Bcl-2与Bax基因在牦牛与黄牛不同组织中的表达差异。结果表明，牦牛Bcl-2编码区全长690 bp，编码229个氨基酸；与黄牛Bcl-2基因核苷酸序列同源性最高，为99.86%，其次是山羊、绵羊，同源性分别为98.41%、97.97%；系统进化树表明，牦牛与黄牛亲缘关系最近。牦牛Bax基因编码区全长579 bp，编码192个氨基酸，与黄牛、藏山羊和金堂黑山羊同源性较高，分别为99.83%、99.48%和99.48%，其次是绵羊、马、人，同源性分别为99.14%、95.34%、94.30%；系统进化树表明，牦牛与黄牛亲缘关系最近。Bcl-2和Bax蛋白不存在信号肽，均为酸性不稳定的疏水蛋白。Bcl-2与Bax基因在黄牛及牦牛下丘脑、垂体、卵巢、输卵管和子宫组织中均有表达，其中牦牛卵巢、子宫中Bcl-2基因表达量分别显著和极显著高于黄牛（P<0.05；P<0.01）；牦牛子宫、输卵管中Bax基因表达量显著高于黄牛（P<0.05），牦牛卵巢中Bcl-2/Bax比值极显著高于黄牛（P<0.01），子宫和垂体中显著高于黄牛（P<0.05）。表明Bcl-2与Bax在动物进化中非常保守且在繁殖活动中起重要作用，牦牛卵巢、子宫、输卵管和垂体中的高表达量可能与牦牛处于极端恶劣环境的细胞抗凋亡作用有关。

关键词: 牦牛; 黄牛; Bcl-2基因; Bax基因; 克隆; 组织表达

Abstract:

The aim of this study was to clarify CDS sequences and expression characteristics of Bcl-2 and Bax genes in the reproductive axis of female yak,in order to provide theoretical basis for investigating the potential role of Bcl-2 and Bax in regulation of yak reproduction.The samples of hypothalamus,pituitary,ovary,oviduct and uterus of female yak and cattle were collected.The yak Bcl-2 and Bax genes were amplified,cloned and analyzed by RT-PCR and bioinformatics methods.Their mRNA expression levels in different tissues were detected by Real-time PCR.The results showed that the Bcl-2 gene coding region of yak was 690 bp,encoding 229 amino acid.The homology of Bcl-2 gene in yak with Bos taurus was the highest (99.86%),and followed by Capra hircus (98.41%) and Ovis aries (97.97%).The phylogenetic tree showed that the closest genetic relationship was found between yak and Bos taurus.The Bax gene coding region of yak was 579 bp,encoding 192 amino acids.The homology of Bax gene in yak with Bos taurus,Tibetan goat and Jintang Black goat were higher,which was 99.83%,99.48% and 99.48%,respectively,and followed by Ovis aries (99.14%),Equus caballus (95.34%) and Homo sapiens (94.30%).The phylogenetic tree showed that the closest genetic relationship was found between yak and Bos taurus.There were no signal peptides in Bcl-2 and Bax proteins,all of which were acidic unstable hydrophobins.Real-time PCR results showed that Bcl-2 and Bax genes were expressed in hypothalamus,pituitary,ovary,oviduct and uterus tissues of yak and cattle.The expression of Bcl-2 gene was significantly higher in yak ovary (P<0.05) and uterus (P<0.01) than that in cattle.The expression of Bax in yak uterus and ovidut was significantly higher than that in cattle (P<0.05).The Bcl-2/Bax ratio in the yak ovary was extremely significantly higher than that in cattle (P<0.01),and that in uterus and pituitary were significantly higher than cattle (P<0.05).These results indicated that Bcl-2 and Bax were relatively conservative in animal evolution.Bcl-2 and Bax played an important role in reproductive activities.The high expression level of them in yak ovary,uterus,oviduct and pituitary might be related to the anti-apoptotic effect of yak in extreme environment.

Key words: yak; cattle; Bcl-2 gene; Bax gene; cloning; tissue expression

中图分类号:

S823.8^＋5

夏忆, 王琴, 何向东, 陈莹, 吉格莫体, 字向东. 牦牛Bcl-2、Bax基因克隆及其在生殖轴上的表达分析[J]. 《中国畜牧兽医》, 2019, 46(7): 1881-1889.

XIA Yi, WANG Qin, HE Xiangdong, CHEN Ying, JIGE Moti, ZI Xiangdong. Cloning and Expression Analysis of Bcl-2 and Bax Genes in Reproductive Axis of Yaks[J]. , 2019, 46(7): 1881-1889.

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