草鱼(♀)×赤眼鳟(♂)杂交F1IGF-Ⅰ和IGF-Ⅱ cDNA克隆、序列分析及表达特征  

cDNA Cloning,Sequence Analysis and Expression Characterization of Insulin-like Growth FactorsⅠandⅡin F1 of Ctenopharyngodon idellus(♀)×Squaliobarbus curriculus(♂)

作　　者：徐莹[1] 王荣华 金生振 赵鑫[1] 王静安[1] 丁春华 罗红[1] 李东放 李耀国[1] 肖调义[1] Xu Ying;Wang Ronghua;Jin Shengzhen;Zhao Xin;Wang Jing'an;Ding Chunhua;Luo Hong;Li Dongfang;Li Yaoguo;Xiao Tiaoyi(Hunan Engineering Research Center for Utilization of Characteristics of Aquatic Resources,Hunan Agricultural University,Changsha,410128; Collaborative Innovation Center for Efficient and Health Production of Fisheries in Hunan Province,Hunan University of Arts and Science,Changde,415000)

机构地区：[1]湖南农业大学,湖南省特色水产资源利用工程技术研究中心,长沙410128 [2]湖南文理学院生命与环境科学学院,水产高效健康生产湖南省协同创新中心,常德415000

出　　处：《基因组学与应用生物学》2020年第4期1478-1489,共12页Genomics and Applied Biology

基　　金：国家自然科学基金青年科学基金项目(31802288);国家自然科学基金面上项目(31572615;31272652);中国博士后科学基金特别资助项目(2018T110833);中国博士后科学基金资助项目(2017M612560)共同资助。

摘　　要：为探究草鱼(♀)×赤眼鳟(♂)杂交F1生长性状形成相关分子机理,本研究通过RACE技术(rapid-amplification of cDNA ends, RACE)克隆了杂交F1个体胰岛素样生长因子-Ⅰ(Insulin-like growth factors-Ⅰ,IGF-Ⅰ)和IGF-Ⅱ的cDNA全长序列,利用荧光定量PCR检测了两个基因的组织表达水平,并比较了杂交F1生长性状大、小两个亚群的基因表达水平差异。结果显示:IGF-Ⅰ和IGF-Ⅱ的cDNA全长分别为824 bp和1 707 bp。IGF-Ⅰ和IGF-Ⅱ蛋白均具有胰岛素样生长因子的典型特征,即包括信号肽和B、C、A、D和E5个功能结构域,并具有6个保守的半胱氨酸。IGF-Ⅰ与IGF-Ⅱ的氨基酸序列同源性较低,仅为26%。IGF-Ⅰ与IGF-Ⅱ在杂交F1下丘脑、垂体、肝脏、脾脏、肾脏、头肾、肠、心脏、皮肤、肌肉、性腺和血液中都有表达,且均在肝脏中表达水平最高,心脏中最低。同池饲养F1的基因表达分析显示,大亚群肝脏、血液和肌肉中IGF-Ⅰ表达水平均极显著高于小亚群对应组织中的表达水平(p<0.01),大亚群肝脏中IGF-Ⅱ表达水平极显著高于小亚群(p<0.01),表明IGF-Ⅰ和IGF-Ⅱ的表达水平与杂交F1的生长性状相关联。实验结果可为深入研究草鱼(♀)与赤眼鳟(♂)杂交F1生长性状的分子调控机制提供科学基础。In order to investigate the molecule mechanism involved in growth trait formation of progeny from Ctenopharyngodon idellus(♀)×Squaliobarbus curriculus(♂),full-length cDNAs of Insulin-like growth factors-Ⅰ(IGF-Ⅰ)and IGF-Ⅱwere cloned by RACE technique.Tissue expression levels of the two genes were detected by qPCR,and the significance of expression level difference between the F1 progeny of largest-subpopulation and smallest-subpopulation was compared.The results showed that full-length cDNAs of IGF-Ⅰand IGF-Ⅱwere 824 bp and 1707 bp,respectively.Typical structure charecteristics of IGFs were observed in the deduced amino acids of the two genes,including the signal peptide and B,C,A,D and E domains,and six conservative cysteine residues.A low sequence identity was found between IGF-Ⅰand IGF-Ⅱ(26%).The IGF-Ⅰand IGF-Ⅱwere widely distributed in tissues of hypothalamus,pituitary,liver,spleen,kidney,head kidney,intestine,heart,skin,muscle,gonad and blood.Both of the two genes were highest expressed in the liver and lowest in the heart.The expression levels of IGF-Ⅰin liver,blood and muscle in the largest-subpopulation were extremely significantly higher than those in the corresponding tissues of smallest-subpopulation(p<0.01),and expression level of IGF-Ⅱin the liver was extremely significantly higher than that of the smallest-subpopulation(p<0.01).The results indic­ate that expression levels of IGF-Ⅰand IGF-Ⅱwere associated with growth traits of F1 progeny.The results may provide a scientific basis for further in-depth study of the molecular regulation mechanism of hybrids.

关 键 词：杂交F1 胰岛素样生长因子 全长序列 组织分布 生长性状 

分 类 号：S917.4[农业科学—水产科学]