黄颡鱼DMRT1基因cDNA全长克隆及其表达分析  

Cloning and expression analysis of DMRT1 gene in Pelteobagrus fulvidraco

作　　者：李林[1,2] 梁宏伟[1,3] 李忠[1,3] 罗相忠[1,3] 张志伟[1,2] 朱媛媛[1,2] 邹桂伟[1,2,3] 

机构地区：[1]农业部淡水生物多样性保护与利用重点开放实验室/中国水产科学研究院长江水产研究所,武汉430223 [2]华中农业大学水产学院,武汉430070 [3]中国水产科学研究院淡水渔业研究中心,无锡214081

出　　处：《华中农业大学学报》2012年第2期220-226,共7页Journal of Huazhong Agricultural University

基　　金：国家"863"计划(2011AA100401);中央级公益性科研院所基本科研业务费专项资金项目

摘　　要：利用RT-PCR和cDNA末端快速扩增技术(rapid amplification of cDNA ends,RACE)克隆了黄颡鱼DMRT1基因cDNA全长序列,并利用实时荧光定量RT-PCR技术对该基因在黄颡鱼成体不同组织及不同发育阶段的表达情况进行研究。结果表明,黄颡鱼DMRT1基因cDNA序列全长1 381bp,其中5′端非翻译区30bp,3′端非翻译区454bp[不包括poly(A)],开放阅读框885bp,编码295个氨基酸。氨基酸序列同源性分析表明,黄颡鱼DMRT1基因与革胡子鲶同源性最高(为81%),与黑鲷、虹鳟、斑马鱼、青鳉的同源性分别为60%、59%、64%和52%,与小鼠、人的同源性较低,分别为42%和44%。实时荧光定量RT-PCR分析表明:DMRT1基因在黄颡鱼胚胎发育阶段及胚后发育的1～51d仔鱼均有表达,且在胚后发育的第31天表达量最高;在成体,只在雄性精巢中特异性表达,其他组织均无表达,且性腺发育阶段的Ⅳ期精巢表达量最高,表明该基因可能在黄颡鱼雄性性腺的形成或功能维持上具有重要作用。The DMRT1 gene was cloned by RT-PCR and rapid amplification of cDNA ends（RACE） methods in yellow catfish （Pelteobagrusfulvidraco）. The expression of the gene was analyzed in adult tissues and different developmental stages by real-time quantitative RT-PCR. Sequence analysis revealed that the full-length of cDNA was 1 381 bp,containing 30 hp 5r-untranslated region,454 bp 3＇-untranslated region [-excluding poly（A）] and 885 bp open reading frame（ORF）,which encode 295 amino acids. A- lignment analysis showed that the amino acid sequences of DMRT1 gene in yellow catfish （Pelteobagrus fulvidraco） were 81% ,60% ,59% ,64% ,52% ,42% and 44% identical to that from clarias lazera（Clarias gariepinus ） , black porgy （ Acanthopagrus schlegelii ）, rainbow trout （ Oncorhynchus m ykiss ）, zebrafish（Danio rerio ）, medaka（Oryzias latipes ）, mouse（Mus musculus ） and human（Homo sapiens ）, respectively. Real-time quantitative PCR analysis indicated that the DMRT1 gene expressed in the whole embryonic development stages and 51-day juveniles of postembryonic development, but the expression reached the highest level in juveniles at 31-day after hatching. During the gonad development stages, it was expressed in the male testis specifically. In the adult fish, the expression was detected only in the male testis, but not in other tissues. These results indicated that the DMRT1 gene may play an important role in the gonad formation or function maintaining of male yellow catfish （Pelteobagrus fulvidraco）.

关 键 词：黄颡鱼 DMRT1基因 cDNA末端快速扩增(RACE) 基因克隆 荧光定量 表达分析 

分 类 号：Q781[生物学—分子生物学] Q786