抚仙金线鲃促性腺激素亚基基因克隆及组织表达分析  

Gonadotropin Subunits cDNA Cloning and Tissue Expression in Sinocyclocheilus tingi

作　　者：杨国坤 关佳佳 孙彩云 王晓爱[2] 潘晓赋[2] 杨君兴[2] 李文笙 

机构地区：[1]有害生物控制与资源利用国家重点实验室,广东省水生经济动物良种繁育重点实验室,中山大学深圳研究院,中山大学生命科学学院,广州510275 [2]遗传资源与进化国家重点实验室,中国科学院昆明动物研究所,昆明650223

出　　处：《四川动物》2016年第5期686-690,共5页Sichuan Journal of Zoology

基　　金：中山大学高校基本科研业务费项目;深圳市战略性新兴产业发展专项资金项目(NYSW20140401010064);云南省应用基础研究面上项目(2012FB183);中国科学院昆明动物研究所“一三五”重大专项(Y206B51181);云南省生物多样性保护专项资金项目

摘　　要：抚仙金线鲃Sinocyclocheilus tingi是云南抚仙湖特有种,虽然实现了人工繁殖,但在塘养环境下仍无法自然繁衍。鱼类的生殖活动受下丘脑-垂体-性腺轴的调控,其中促性腺激素在该过程中起重要作用。本实验利用实时荧光定量PCR和c DNA末端快速扩增技术从抚仙金线鲃垂体中克隆了GTHα、FSHβ和LHβ3种基因的c DNA序列,其中GTHα的开放阅读框(ORF)长度为357 bp,编码118个氨基酸,有10个半胱氨酸残基和2个N-糖基化位点;FSHβc DNA全长为856 bp,ORF长度为393 bp,编码130个氨基酸,有11个半胱氨酸残基和1个N-糖基化位点;LHβc DNA全长为930 bp,ORF长度为441 bp,编码146个氨基酸,有12个半胱氨酸残基和1个N-糖基化位点。实时荧光定量PCR结果显示,GTHα和LHβ都只在雄鱼和雌鱼的垂体中表达。FSHβ在垂体中表达量最高,在雌鱼和雄鱼的脂肪、肌肉和雄鱼的精巢和肝脏中有少量表达。本研究为抚仙金线鲃的人工繁育提供了重要的基础数据。Sinocyclocheilus tingi is an endemic species of the Fuxian Lake in Yunnan province. Although the artificial breeding has been successfully developed,the natural reproduction of S. tingi is still failed in pond culture. It is well known that fish reproduction is regulated by the hypothalamus-pituitary-gonadal axis,and the gonadotropins（ GTHs） play important role in this process. In this study,the sequences of GTHα,FSHβ and LHβ were obtained from the pituitary of S. tingi using real-time quantitative PCR（ q PCR） and rapid amplification of c DNA ends technology. The results showed that the open reading frame of GTHα was 357 bp,encoding 118 amino acids,containing 10 cysteine knot motifs and 2 N-linked glycosylation sites. The full length of FSHβ c DNA was 856 bp,with 393 bp open reading frame in length encoding the proteins of 130 amino acids,containing 11 cysteine knot motifs and a N-linked glycosylation site. The full length of LHβ c DNA was930 bp,with 441 bp open reading frame in length encoding proteins of 146 amino acids,containing 12 cysteine knot motifs and a N-linked glycosylation site. The result of q PCR indicated that GTHα and LHβ were only expressed in the pituitary of both male and female fish. The highest expression level of FSHβ was observed in the pituitary,and the even lower levels were detected in the fat tissue,muscle of male and female fish,and testis and liver of male fish. These findings provided basic data for further research in the reproduction of S. tingi.

关 键 词：抚仙金线鲃 促性腺激素 克隆 组织表达 

分 类 号：S917.4[农业科学—水产科学]