牦牛STAG3基因克隆及其在不同年龄段牦牛睾丸中的表达定位
Clone of STAG3 Gene and Its Expression and Localization in Yak(Bos grunniens)Testis with Different Ages
石军 1陈文丽 1白旭 1李建富 1袁宝 1赵兴绪 2张全伟2
作者信息
- 1. 甘肃农业大学生命科学技术学院,兰州 730070;甘肃省动物生殖生理及繁殖调控重点实验室,兰州 730070
- 2. 甘肃农业大学生命科学技术学院,兰州 730070;甘肃农业大学动物医学院,兰州 730070;甘肃省动物生殖生理及繁殖调控重点实验室,兰州 730070
- 折叠
摘要
基质抗原3(stromal antigen 3,STAG3)是黏连蛋白复合体的重要成分之一,在动物卵泡发育、卵巢早衰和癌症发生等生物学过程中发挥重要调节作用,其在牦牛(Bos grunniens)生殖和发育过程中的作用尚不明确.本研究以不同岁龄(2,4,6和8岁龄)牦牛睾丸组织为材料,通过反转录PCR(reverse transcription-PCR,RT-PCR)克隆STAG3 基因编码区序列(CDS),利用STRING蛋白质互作数据库(STRING:functional protein association networks)和基因本体(Gene Ontology,GO)预测分析STAG3互作蛋白及生物学功能;通过苏木精-伊红(hematoxylin-eosin,HE)染色观察不同年龄牦牛睾丸组织形态结构;免疫组织化学(immunohistochemistry,IHC)染色和qRT-PCR分析STAG3分布及表达规律.结果显示,牦牛STAG3基因CDS区全长3679 bp,开放阅读框为1659 bp,编码552个氨基酸,蛋白无跨膜结构区,可能参与第一次减数分裂、联会复合和染色体浓缩等生物学过程;STAG3蛋白在各年龄段牦牛睾丸组织中均有表达,主要定位于初级精母细胞和次级精母细胞;2岁龄牦牛睾丸组织STAG3表达最高,且随年龄增长表达逐渐降低.结果表明,STAG3在动物进化中较为保守,各年龄段牦牛睾丸组织均有表达,其可能在牦牛生殖过程中发挥重要作用.本研究为解析牦牛生殖过程中STAG3功能及调控机制提供了参考依据.
Abstract
Stromal antigen 3(STAG3),one component of adhesin complexes,plays important role in regulation of follicular development,premature ovarian failure and carcinogenesis in animals.However,its function and regulatory mechanisms are incompletely understood in yak(Bos grunniens).In the present study,the yak testicular tissues with different ages(2,4,6 and 8 years old)were collected.The coding sequence(CDS)of STAG3 gene was cloned using reverse transcription-PCR(RT-PCR).The interacted proteins and biological functions of STAG3 were predicted using STRING(functional protein association networks)and Gene Ontology(GO)procedures.Histomorphological structure of yak tissues was observed using hematoxylin-eosin(HE)staining.The localization and expression pattern analysis of STAG3 were evaluated using immunohistochemical(IHC)staining and qRT-PCR.The results showed that the CDS of yak STAG3 was 3679 bp with a 1659 bp open reading frame,and encoding 552 amino acids without transmembrane structure region.STAG3 might involve in the meiosis Ⅰ phase,synaptic re-combination and chromosome concentration.The STAG3 protein was widely expressed in testicular tissues,and were located mainly in primary spermatocyte and secondary spermatocyte.The highest expression level of STAG3 was presented in the yak testis of 2 years old,and was gradually decreased with ages.In conclusion,STAG3 was conservative in animal evolution,and widely expressed in the yak testises,which might play an important role in yak reproductive processes.This study provides basis for uncovering the function and mechanism of STAG3 in yak reproduction.
关键词
牦牛/基质抗原3(STAG3)/基因克隆/生物信息学分析Key words
Yak/Stromal antigen 3(STAG3)/Gene cloning/Bioinformatics analysis引用本文复制引用
基金项目
国家自然科学基金(03120057)
中国科学院"西部之光"人才培养计划(03321013)
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据