异齿裂腹鱼Clock基因cDNA的克隆与组织表达特征  

Cloning and Expression Profile of Clock Gene cDNA in Schizothorax o′connori

作　　者：周建设 扎西拉姆[1] 王万良 孙帅杰 张驰 ZHOU Jianshe;TASHI Lhamo;WANG Wanliang;SUN Shuaijie;ZHANG Chi(Institute of Fisheries Science,Xizang Academy of Agricultural and Animal Husbandry Sciences,Lhasa 850032,China;College of Animal Science and Technology,Henan Agricultural University,Zhengzhou 450002,China)

机构地区：[1]西藏自治区农牧科学院水产科学研究所,西藏拉萨850032 [2]河南农业大学动物科技学院,河南郑州450002

出　　处：《水产科学》2023年第1期48-56,共9页Fisheries Science

基　　金：西藏自治区自然科学基金资助项目(XZ2019ZRG-103)。

摘　　要：异齿裂腹鱼广布于雅鲁藏布江,产卵期为每年的3—5月,表现出明显的季节性繁殖行为,Clock基因被认为是自然选择塑造日节律以及与动物繁殖等生活史特性有关潜在的目标基因,异齿裂腹鱼Clock蛋白C末端Poly-Q长度的变异可能与其产卵开始的时间关联。以采自雅鲁藏布江的异齿裂腹鱼为对象,采用反转录PCR和cDNA末端快速扩增技术克隆异齿裂腹鱼Clock基因cDNA全长序列,通过实时荧光定量PCR对其在异齿裂腹鱼雌、雄个体不同组织中的表达特征进行分析。试验结果显示:异齿裂腹鱼Clock基因cDNA全长为4291 bp(Genbank登录号MT774361),5′非编码区为539 bp,3′非编码区为1046 bp,编码序列为2706 bp,共编码901个氨基酸,所编码的氨基酸在末端具有典型的Poly-Q结构;Clock基因在异齿裂腹鱼雌、雄个体的肝脏、脾脏、心脏、脑、性腺和肌肉中均有表达,且以性腺中的相对表达量最高,肌肉次之,脾脏最低;Clock基因除在心脏组织中的相对表达量雄鱼高于雌鱼,在其他组织的相对表达量均表现为雌鱼高于雄鱼。异齿裂腹鱼雌、雄个体的Clock基因在性腺组织中均高表达,表明Clock基因在异齿裂腹鱼的繁殖过程中发挥着更大的作用,异齿裂腹鱼Clock蛋白大片段的插入也可能是为了适应高海拔某些条件而特化的。Schizothorax o′connori is widely distributed in the Brahmaputra River and spawns from March to May,showing obvious seasonal breeding behavior.The Clock gene was considered as a potential target gene for natural selection to shape the daily rhythm and to be related to life history characteristics such as animal reproduction.The variation in the length of C-terminal Poly-Q of Clock protein of S.o′connori may be involved in the onset of oviposition.In order to establish the foundation for understanding of the important role of Clock gene of Schizothorax in Qinghai-Xizang Plateau in seasonal breeding behavior,the complete cDNA sequence of Clock gene in S.o′connori were cloned by RT-PCR and RACE-PCR,and the expression profile were detected in liver,spleen,heart,brain,gonad and muscle of both male and female.The results showed that the complete cDNA sequence of Clock gene was 4291 bp(GenBank:MT774361),5′UTR was 539 bp,3′UTR was 1046 bp,CDS was 2706 bp,and encoded 901 amino acid,with typical Poly-Q structure in the protein sequence.The expression profile of Clock gene was existed in all detected tissue of S.o′connori,with the maximum in gonad and the minimum in spleen.The expression profile of all detected tissue revealed a trend that higher expression profile in female than that in male,except heart.The high expression profile of Clock gene in both female and male gonad illustrated that more active important role in reproduction process of S.o′connori,and the insertion of large fragments of Clock protein may also be specialized to accommodate certain conditions at high altitudes.

关 键 词：异齿裂腹鱼 CLOCK基因 克隆 组织表达 

分 类 号：Q786[生物学—分子生物学]