沼泽型水牛FST基因克隆分析与组织表达模式  

Cloning and expression pattern investigation of swamp buffalo FST gene

作　　者：汪浩鑫 陈维丽 张俊[1] 许建春 自永宏 杨小芬 石德顺[1] 陆凤花[1] WANG Haoxin;CHEN Weili;ZHANG Jun;XU Jianchun;ZI Yonghong;YANG Xiaofen;SHI Deshun;LU Fenghua(State Key Laboratory of Conservation and Uhilization of Subtropical Agror Bioresources,Guangxi University,Narning 530005,China;Key Laboratory of Animal Biotechnology.Minisry of Agriculture,Norihwest A&F Universily,Yangling,Shanxi 712100,China)

机构地区：[1]广西大学亚热带农业生物资源保护与利用国家重点实验室,广西南宁530005 [2]西北农林科技大学农业部动物生物技术重点实验室,陕西杨凌712100

出　　处：《中国兽医学报》2022年第9期1909-1914,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31760666);广西自然科学基金资助项目(2020JJD130069)

摘　　要：旨在克隆水牛卵泡抑素(follistatin, FST)基因,并对该基因进行生物信息学分析,检测其在水牛不同组织中的表达情况,为进一步探讨FST在水牛胚胎发育过程中的功能奠定基础。从水牛卵巢组织中提取RNA,反转录成cDNA后,利用RT-PCR技术克隆获得水牛FST基因CDS区的全长序列,并进行生物信息学分析。结果显示,水牛FST基因编码区全长1 035 bp,编码344个氨基酸。水牛FST基因与黄牛、鸡、绵羊、人、黑猩猩、褐家鼠、野猪和山羊的同源性分别为99.1%,81.4%,98.4%,91.9%,91.4%,89.3%,93.4%和81.4%,且在不同物种间高度保守,符合生物的进化规律。FST蛋白包含40个α-螺旋、63个延伸链及241个无规则卷曲,分别占11.6%(40个),18.3%(63个)和70.0%(241个),为经典分泌蛋白。qRT-PCR结果显示,FST在水牛胃、肌肉、卵巢、睾丸、心脏、肝脏、肺脏和脾脏组织中均有表达,其中在肺脏和卵巢中表达最高,睾丸中表达最低。本研究克隆得到了沼泽型水牛FST基因,并对不同组织内FST进行了基础的生物信息学分析,为阐明FST在水牛胚胎发育过程中的功能,提高水牛胚胎发育水平提供理论基础。The aim of this study was to clone buffalo follistatin(FST) gene.Bioinformatics analysis of FST gene was conducted to detect its expression in different tissues of buffaloes, laying a foundation for further study on the function of FST in embryonic development of buffaloes.After RNA was extracted from buffalo ovarian tissue and reversely transcribed into cDNA,the full-length sequence of buffalo FST gene CDS region was cloned by RT-PCR and analyzed by bioinformatics.The results showed that the coding region of buffalo FST gene was 1 035 bp in length, encoding 344 amino acids.The homology of buffalo FST gene with bovine, chicken, sheep, human, chimpanzee, Rattus norvegicus, wild boar and goat was 99.1%,81.4%,98.4%,91.9%,91.4%,89.3%,93.4% and 81.4%,respectively.They are highly conserved among different species and conform to the law of biological evolution.FST protein contains 40 α-helices, 63 extended strands and 241 random coils, accounting for 11.6%(40),18.3%(63) and 70.0%(241) of classical secreted proteins, respectively.qRT-PCR results showed that FST was expressed in buffalo stomach, muscle, ovary, testicular heart, liver, lung and spleen, with the highest expression in lung and ovary, and the lowest expression in testicular.In this study,FST genes of swamp buffalo were cloned,and basic bioinformatics analysis of FST in different tissues was conducted to provide theoretical basis for elucidating the function of FST in the development of buffalo embryos and improving the development level of buffalo embryos.

关 键 词：水牛 FST 卵母细胞体外成熟 基因克隆 胚胎发育 

分 类 号：S823.83[农业科学—畜牧学]