圆斑星鲽sox9基因的克隆与表达  

Cloning and Expression of the sox9 Gene in Spotted Halibut(Verasper variegatus)

作　　者：张乐乐 边力[2] 常青[2] 侯吉伦[3] 陈四清[2] 赵庆[1,2] 刘琨 葛建龙[2] 刘长琳[2] ZHANG Lele;BIAN Li;CHANG Qing;HOU Jilun;CHEN Siqing;ZHAO Qing;LIU Kun;GE Jianlong;LIU Changlin(National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai201306;Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao266071;Beidaihe Central Experiment Station,Chinese Academy of Fishery Sciences,Qinhuangdao066100)

机构地区：[1]水产科学国家级实验教学示范中心,上海海洋大学,上海201306 [2]中国水产科学研究院黄海水产研究所,青岛266071 [3]中国水产科学研究院北戴河中心实验站,秦皇岛066100

出　　处：《渔业科学进展》2018年第6期72-80,共9页Progress in Fishery Sciences

基　　金：中国水产科学研究院黄海水产研究所基本科研业务费(20603022016005);天津市农业科技成果转化与推广项目(201604100)共同资助.

摘　　要：本研究筛选到圆斑星鲽(Verasper variegatus)的性别相关基因sox9,并通过RACE技术获得了全长序列,基因全长为3287 bp,包括1431 bp的ORF,编码477个氨基酸,368 bp的5'UTR和1488 bp的3'UTR。在3'UTR中有多聚腺苷酸尾和加尾信号AATAAA。通过荧光定量PCR测定了sox9基因在圆斑星鲽成鱼不同组织中的表达水平,发现sox9基因在圆斑星鲽的脑、眼、鳃、心、肝、胆、肠、精巢、卵巢、肾和肌肉等各个组织中都有不同程度的表达。在鳃、脑和精巢组织中检测到较高水平的sox9转录,其中精巢中的转录水平显著高于其他组织,sox9基因在性腺中的表达显示出性别两相性差异。其在精巢中的表达水平要显著高于卵巢,说明sox9基因与雄性性腺发育相关。通过测定sox9基因在圆斑星鲽幼鱼不同发育时期(20、30、40、50、60、70和80日龄)的表达水平,发现其在20～50日龄表达量逐渐下降,在60日龄时表达量上升,推测表达量上升可能与幼鱼性腺分化相关。Significant differences exist in the growth rate and body size of male and female spotted halibut(Verasper variegatus),with the females being significantly larger than the males.All-female cultivation can improve breeding efficiency.Studies investigating the mechanism of sex determination and features of sex differentiation may provide valuable information for all-female cultivation.In this study,we successfully identified the sox9gene of spotted halibut.The total length is3287bp,including a 1431bp ORF,encoding477amino acids;the5'UTR is368bp and the3'UTR is1488bp.Polyadenylic acid tails and AATAAA caudal signals were identified in the3'UTR.The expression of the sox9gene differed in the brain,eye,gills,heart,liver,gallbladder,intestine,testis,ovary,kidney,and muscle in spotted halibut.Transcript levels were high in the gill,brain,and testis,and were significantly higher in the testis than in the other tissues.A two-phase gender difference was observed in the gonads;sox9 expression was significantly higher in the testis than in the ovary,indicating that sox9might be associated with male gonad development.The dynamic expression patterns of the sox9gene in spotted halibut larvae gradually declined from20to50d after hatching,with a sudden rise observed after60d.This increased expression might be associated with differentiation of larval gonad.

关 键 词：圆斑星鲽 SOX9 性别基因 MRNA表达 

分 类 号：S917.4[农业科学—水产科学]