双棘黄姑鱼IGF3基因克隆及表达模式分析  

IGF3 gene cloning and expression pattern of Protonibea diacanthus

作　　者：林权卓[1] 陈奕彬[1] 胡娟[1] 王树启[2] 杨宪宽 沈卓坤 赵会宏[1] 

机构地区：[1]华南农业大学动物科学学院,广东广州510642 [2]汕头大学广东省海洋生物技术重点实验室,广东汕头515063 [3]广东省海洋工程职业技术学校,广东广州510320

出　　处：《南方水产科学》2016年第1期50-58,共9页South China Fisheries Science

基　　金：国家科技基础平台建设项目(2004DKA30470-010);公益性行业(农业)科研专项经费项目(201303048);广东省科技计划项目(2012B040400004);广东省海洋生物技术重点实验室开放基金(GPKLMB201301);广东省海洋渔业科技推广专项(A201201E04)

摘　　要：以双棘黄姑鱼(Protonibea diacanthus)为实验对象,通过RACE方法克隆得到IGFs家族成员IGF3。双棘黄姑鱼IGF3 c DNA全长1 184 bp。氨基酸一致性分析显示,双棘黄姑鱼IGF3与尼罗罗非鱼(Oreochromis niloticus)一致性最高,仅为58.0%;与自身IGF1的2个亚型及IGF2相比,一致性分别低至26.2%、25.2%和21.9%。系统进化树显示鱼类IGF3基因先与IGF2聚类,再与IGF1汇聚构成IGF家族,最后IGFs与胰岛素聚类。组织表达模式显示IGF3在双棘黄姑鱼心脏、性腺、脑区均有表达,在性腺中表达最强。实时定量PCR结果表明,IGF3在精巢发育期表达量显著高于其他时期,呈显著下降趋势;在卵巢成熟期的表达量显著高于其他时期。此外,雌鱼在成熟期前、中、后脑IGF3的表达量显著高于其他时期,而雄鱼各脑区IGF3表达趋势呈多样性。上述结果表明,IGF3可能参与了双棘黄姑鱼精巢发育和卵巢成熟期生殖功能的调节,且前脑可能主要参与了IGF3在性腺表达的反馈调节。We cloned IGF-Ⅲ gene from Protonibea diacanthus by RACE method. The length of c DNA was 1 184 bp. The homologous analysis of deduced protein shows that the identity of IGF-Ⅲ gene and Oreochromis niloticus was 58. 0%. Compared with IGF-Ⅱ and two subtypes of IGF-Ⅰ of P. diacantus,the identities were 21. 9%,26. 2% and 25. 2%,respectively. Phylogenetic tree analysis shows that IGF-Ⅲ tended to cluster with IGF-Ⅱ firstly and then cluster with IGF-Ⅰ composing IGF family,and finally IGF family clustered in the same branch with Insulin. Tissue distribution shows that IGF-Ⅲ was widely expressed in heart,gonad and brain of P. diacanthus,especially highest in gonad. Quantitative real-time PCR analysis reveals that the expression of IGF-Ⅲ in testis was significantly higher in developing phase than that in the others,showing a significant decline. The expression of IGF-Ⅲ in ovary of maturation phase was also remarkably higher than that in other periods. Besides,the expression of IGF-Ⅲ in forebrain,midbrain and hindbrain in female fish in mature phase was higher than that in other periods. However,IGF-Ⅲ expression pattern was different in malefish. It is suggested that IGF-Ⅲ might be important for the development of testis and maturation of ovary,and forebrain might involve in the feedback regulation of IGF-Ⅲ expression in gonad.

关 键 词：双棘黄姑鱼 IGF3 基因克隆 表达模式 

分 类 号：S917.4[农业科学—水产科学]