小体鲟AR基因克隆、序列分析及其组织mRNA表达  

Cloning, sequence analysis, and mRNA expression distribution of the androgen receptor from Acipense ruthenus

作　　者：余晶[1,2] 董晓丽[1] 张颖[1] 刘晓勇 潘鹏 买丽坎 孙大江[1] 

机构地区：[1]中国水产科学研究院黑龙江水产研究所,黑龙江哈尔滨150070 [2]上海海洋大学水产与生命学院,上海201306 [3]中国水产研究院鲟鱼繁育工程中心,北京100039

出　　处：《中国水产科学》2014年第6期1146-1153,共8页Journal of Fishery Sciences of China

基　　金：国家公益性行业(农业)科研专项(201003055);国家重大科技成果转化项目(ZD-2012-345);国家科技支撑计划项目(2012BAD25B10)

摘　　要：采用RT-PCR和RACE技术,克隆了小体鲟(Acipenser ruthenus)雄激素受体(AR)基因c DNA全长序列,应用real-time q PCR法对雌、雄个体不同组织中AR m RNA的表达进行检测。序列分析表明,AR基因c DNA全长为2 953 bp,包括5′端68 bp的非编码区(untranslation region,UTR),2 528 bp的开放阅读框ORF(open reading frame)和3′端327 bp的非编码区(不包括ploy A的尾巴)。AR前体蛋白由843个氨基酸组成,理论分子质量为94.24 k D。同源比较结果表明,小体鲟AR氨基酸序列与其他鱼类的AR氨基酸序列同源性较高,与杂交鲟(A.ruthenus×Huso huso)、西伯利亚鲟(A.baerii)、虹鳟(Oncorhynchus mykiss)和日本鳗鲡(Anguilla japonica)的雄激素受体同源性分别为99%、98%、83%和83%。real-time q PCR分析小体鲟雌雄个体不同组织中的表达结果显示,AR基因在雌雄个体中相对表达基本相同,都是在性腺、肌肉和肾中高量表达,而在其他组织中少量表达。AR基因在小体鲟体内广泛表达,表明该基因可能对其生长和繁殖有重要作用。Sterlet （Acipenser ruthenus） belongs to the Acipenseridae which is one of the oldest families of fishes. Sterlet are suitable for large scale aquaculture as they will breed in captivity. Despite their relatively small body size （maxi-mum body length 1 m and mass of 20 kg）, starlet are of great interest to fishermen, fish farmers, and consumers due to the excellent taste of their meat and caviar. Androgen receptor （AR）, a ligand-dependent transcription factor of andro-gens, is important for sexual differentiation, gonadal maturation, reproductive behavior, and maintenance of male sexual characteristics. This study on AR focused on the sequencing and characterization of cDNA encoding, and the expression of AR mRNA in different tissues in A. ruthenus. The full length of AR cDNA was cloned from gonadal tissue by RT-PCR and RACE. AR mRNA expression was detected using real-time qPCR in different male and female tissues. Sequence analysis showed that the AR was 2 953 bp in length, including a 68 bp 5′terminal untranslation region（UTR）, a 2 528 bp encoding region, and a 327 bp 3′terminal UTR（except ploy A）. AR precursor protein was encoded by 843 amino acids and had a predicted molecular weight of 94.24 kD. Alignment analysis showed that the sequence was 99%, 98%, 83%, 83%and 81%similar to those of A. ruthenus×Husohuso, A. baerii, Oncorhynchus mykiss, Anguilla japonica and Ovisaris, respectively, and the coding sequence had a higher homology with fish. Several domains present in all cloned ARs were identified in starlet AR. The domains corresponded to an amino-terminal hypervariable transcriptional activation domain （TAD）, a central highly conserved DNA-binding domain（DBD）, and a carboxy-terminal li-gand-binding domain（LBD）. Percentages of homology-similarity among these functional domains in teleost fish ranged between 60%and 80%for the LBD, 70%and 99%for the DBD, and 10%and 60%for the TAD when compared with those of sterlet. Application of real-time qPCR found that AR mRNA was expre

关 键 词：小体鲟 雄激素受体 基因克隆 序列分析 组织表达 

分 类 号：S917[农业科学—水产科学]