七彩神仙鱼Vasa基因cDNA的克隆及表达分析  

Cloning and expression analysis of Vasa gene in Symphysodon haraldi

作　　者：林睿涓 高建忠[1] 金仕容 赵玉明[1] 陈再忠[1] 

机构地区：[1]上海海洋大学水产与生命学院,上海201306

出　　处：《上海海洋大学学报》2017年第3期330-338,共9页Journal of Shanghai Ocean University

基　　金：上海市农委科技兴农重点攻关项目[沪农科种字(2015)第19号];上海海洋大学动物遗传育种协同创新中心项目(ZF1206)

摘　　要：源自DEAD-box家族的Vasa基因是生殖细胞的分子标记之一。本研究克隆了七彩神仙鱼(Symphysodon haraldi)Vasa基因的全长序列,并进行了不同组织的表达分析。结果表明:七彩神仙鱼Vasa基因c DNA序列共2 370 bp,其中5'UTR占123 bp,3'UTR为279 bp,ORF编码655个氨基酸,长1 968 bp。氨基酸序列同源性分析表明七彩神仙鱼的Vasa基因与尼罗罗非鱼(Oreochromis niloticus)的同源性最高。半定量分析表明,Vasa基因在成熟七彩神仙鱼的性腺中特异表达,在其它组织中无表达信号。qRT-PCR结果显示,Vasa在七彩神仙鱼早期胚胎发育阶段及出膜后50日内均有表达。在受精卵至囊胚期阶段,Vasa基因表达持续增加,在囊胚期至孵化期阶段,表达持续降低。在仔鱼阶段,Vasa分别在25日龄和40日龄出现了最低和最高表达量。繁殖前后比较发现,繁殖前的精巢组织中Vasa表达量比繁殖后的表达量低,而卵巢恰好相反。本研究结果可为研究七彩神仙鱼的性分化、生殖细胞分子标记及其发育提供参考。As a member of DEAD-box family,Vasa is considered to be one of the molecular markers of germ cells. In this study,full length of Vasa c DNA in discus fish（ Symphysodon haraldi） was cloned and expression in different tissues was analyzed. The results showed that the full-length of c DNA was 2 370 bp,containing a 123 bp 5＇-untranslated region,a 279 bp 3＇-untranslated region and a 1 968 bp open reading frame,encoding 655 amino acids. Alignment analysis indicated that the amino acid sequence of Vasa gene in discus fish was most identical with tilapia（ Oreochromis niloticus）. PCR analysis showed clearly that the Vasa gene was especially expressed in the gonads of mature fish,while it was not expressed in the other tissues.qRT-PCR analysis indicated that the Vasa gene was expressed from the whole embryonic developmental stages to the first 50-days juveniles. The Vasa gene expression increased continuously from fertilized eggs to blastula stage,then decreased from blastula stage to hatching. At larval stages,Vasa exhibited the lowest and highest expression on 25 dph（ days post hatching） and 40 dph,respectively. By comparison,the expression in testis increased after reproduction which is contrast to the ovary. The findings in this study can provide information to study sexual differentiation,molecular markers and development of germ cell in discus fish.

关 键 词：七彩神仙鱼 VASA基因 克隆 表达分析 

分 类 号：S917[农业科学—水产科学]