半滑舌鳎DMRT 1基因的克隆与表达分析

半滑舌鳎DMRT 1基因的克隆与表达分析 Cloning and Expression Analysis of DMRT 1 Gene in Cynoglossus semilaevis 孙业盈 SUN Yeying 1 2 first-author 张全启 ZHANG Quanqi 1 2 齐洁 QI Jie 1 2 王志刚 WANG Zhigang 1 2 陈妍婕 CHEN Yanjie 1 2 李春梅 LI Chunmei 1 2 钟其旺 ZHONG Qiwang 1 2 中国海洋大学海洋生命学院 中国海洋大学海洋生命学院 College of Marine Life Science,Ocean University of China College of Marine Life Science,Ocean University of China 中国海洋大学海洋生命学院,山东,青岛,266003 中国海洋大学海洋生命学院,山东,青岛,266003 利用同源克隆的方法克隆了半滑舌鳎（Cynoglossus semilaevis）的DMRT 1基因;并用RT-PCR技术检测了该基因在半滑舌鳎不同发育时期以及雌雄成鱼的不同组织的表达情况.结果表明,该基因cDNA序列全长1 232 bp（GenBank登录号为EU007655）,包括774 bp开放阅读框,131 bp5′非翻译区以及含poly（A）信号的329 bp的3′非翻译区,编码258个氨基酸.氨基酸序列同源性分析表明,半滑舌鳎DMRT1氨基酸序列与牙银汉鱼、黑鲷、青鳉、河豚和斑马鱼DMRT1氨基酸序列的同源性分别达到了63%、59%、54%、53%和52%;与光滑爪蟾、小鼠和人类的同源性较低分别为38%、42%和41%.RT-PCR分析表明,DMRT 1基因在半滑舌鳎早期胚胎不同发育时期和孵化后5、13、18、22和35 d的仔鱼均有表达,在孵化后22 d表达量最高.在成体鱼中只在雄性精巢中有特异表达,其他组织均无表达,表明该基因可能参与半滑舌鳎雄性性腺的发育或精子的形成. The DMRT1 gene was obtained by homologous cloning in Cynoglossus semilaevis.The expression of the gene was analyzed in adult tissues and different developmental stages by RT-PCR.Sequence analysis revealed that the full-length cDNA was 1 232 bp containing 774 bp open reading frame（ORF）, 131 bp 5′UTR,329bp 3′UTR.This sequence encoded 258 amino acids.Alignment analysis showed the amino acid sequence was 63%,59%,54%,53%,52%,38%,42%,41% identical to that from pejerrey,black sea bream,medaka,fugu,zebrafish,Africa clawed frogs,mouse and human,respectively.RT-PCR analysis showed that the gene expressed in all the analyzed developmental stages,from early embryo to 35-day juveniles,but the expression reached the highest level in juveniles at 22-days after hatching.In adult fish,however,its expression was detected only in male testis.This result indicated that the DMRT1 may play important role in the gonad differentiation and maturation of male C.semilaevis. 半滑舌鳎 DMRT1基因 克隆 表达分析 Cynoglossus semilaevis DMRT1 gene cloning expression analysis Q78 山东省良种工程重大课题 2008-02-01 2021-04-01 2