海产花鲈IGFBP-1、2基因的克隆及表达分析  

Cloning and Characteristics of Insulin-Like Growth Factor-Binding Protein-1、2 Genes(IGFBP-1、2) of Japanese Sea Bass(Lateolabrax japonicus) and Their Expression Analysis

作　　者：钱焜[1] 温海深[1] 迟美丽[1] 倪蒙[1] 张冬茜[1] 丁玉霞[1] 

机构地区：[1]中国海洋大学海水养殖教育部重点实验室,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2014年第9期37-45,共9页Periodical of Ocean University of China

基　　金："十二五"国家科技支撑计划重大项目(2011BAD13B03)资助

摘　　要：利用逆转录聚合酶链式反应(RT-PCR)和cDNA末端快速扩增法(RACE)克隆得到花鲈的IGFBP-1和IGFBP-2基因cDNA全长序列,IGFBP-1全长1 779bp,编码248个氨基酸,IGFBP-2全长815bp,编码267个氨基酸。花鲈的IGFBP-1和IGFBP-2氨基酸序列同其他硬骨鱼类一样都包括C-端结构域、N-端结构域和多变的中央L-结构域。二者均存在18个保守的半胱氨酸残基以维持氨基酸二级结构的稳定,同时也都拥有N-端结构域的CGCCXXC-box和C-端结构域的CWCV-box,它们都是与IGF配体结合的重要结构。此外,IGFBP-2的C-端结构域还存在一个RGD结构,它能够与整合素相互作用,介导细胞与细胞之间、细胞与细胞外基质之间的黏附作用。利用荧光实时定量PCR技术对花鲈的脑、垂体、心脏、鳃、胃、盲肠、肠、头肾、肾脏、肝脏、性腺(雄)、脾脏和肌肉这13个组织的IGFBP-1和IGFBP-2mRNA的相对表达水平进行分析。结果显示,花鲈IGFBP-1mRNA表达量较为广泛,在肝脏中的转录水平最高,在肌肉和脾脏有较多的表达。花鲈IGFBP-2mRNA仅在肝脏中的表达水平最高,在肌肉中表达水平较高,其他组织表达量较低。RT-PCR and rapid amplification of cDNA ends（RACE）were used to isolate the full length cDNA sequence of IGFBP-1and IGFBP-2from Japanese sea bass（Lateolabrax japonicus）.The full-length cDNA of IGFBP-1was 1 779 bp in length,encoding 248 amino acids and the full-length cDNA of IGFBP-2was815 bp in length,encoding 267 amino acids.The proteins deduced from IGFBP-1and IGFBP-2contained a C-terminal and a N-terminal domain and a variable central L-domain as other teleosts.In addition,two proteins contained 18 conserved cysteine residues,CGCCXXC-box at N-terminal and CWCV-box at C-terminal,which may contribute to their stability or associate with their ligands.The RGD motif at C-terminal of IGFBP-2was shown to interact with integrin,a family of cell adhesion receptors involved in cell-to-cell interaction and intracellular matrix association.The abundance of IGFBP-1and IGFBP-2transcript in different tissues was determined with florescent real-time quantitative RT-PCR.The highest abundance of IGFBP-1transcript was detected in liver,which was followed by muscle and spleen.The highest abundance of IGFBP-2transcript was also detected in liver which was followed by muscle.

关 键 词：花鲈 类胰岛素生长因子结合蛋白-1 类胰岛素生长因子结合蛋白-2 荧光实时定量PCR 组织表达 

分 类 号：S917.4[农业科学—水产科学]