黄颡鱼胰岛素样生长因子结合蛋白1基因的克隆及表达分析

Expression and expression analysis of IGFBP1 gene in yellow catfish(Pelteobagrus fulvidraco)

为研究黄颡鱼(Pelteobagrus fulvidraco)胰岛素样生长因子结合蛋白1基因(IGFBP1)的特征,阐明IGFBP1基因在不同性别黄颡鱼组织中的表达特征和IGFBP1蛋白在雌、雄黄颡鱼肌肉中的表达差异,克隆IGFBP1基因的c DNA全长,并进行实时荧光定量PCR检测和Western Blot分析。结果表明:黄颡鱼IGFBP1基因c DNA全长为1 259 bp,其开放阅读框长度为714 bp,编码237个氨基酸,3'端和5'端非翻译区分别为407 bp和138 bp;黄颡鱼IGFBP1基因核苷酸序列与鲇形目斑点叉尾鮰的相似度(89%)较高,与鲢、鲤、鲫、草鱼和团头鲂等的差异较大;肝脏是黄颡鱼IGFBP1基因m RNA表达最丰富的组织,IGFBP1基因在雄性个体心脏、肾脏、肌肉和肠组织中的表达量显著高于雌性的(P<0.05);IGFBP1蛋白在雌、雄黄颡鱼肌肉组织中都有表达,且在雄性个体中的表达量高于在雌性个体中的表达量。

To analyze the characteristics of insulin-like growth factor-binding protein I（IGFBP1）, and to reveal its expression pattern in different gender tissues and the expression difference of its protein between male and female muscles, the 1GFBP1 cDNA was cloned from the Pelteobagrus fulvidraco. The mRNA expression of IGFBP1 was studied using florescent quantitative real-time PCR and the protein expression pattern of IGFBP1 was detected using western blot method. The full-length of cDNA of lGFBP1 is 1 259 bp which included a 138 bp complete 5＇untranslated region （5＇ UTR）, a 407 bp 3＇ UTR and a 714 bp open reading frame （ORF） region which encoded a 237 amino acid of peptides. The results indicated that nucleotide sequences of 1GFBP1 were obvious difference from other species （such as Hypophthalmichthys molitrix, Cyprinus carpio, Carassius auratus, Ctenopharynodon idellus and Megalobrama amblycephala, etc.） except for Ictalurus punctatus, which was identical to Siluriformes of Pelteobagrus fulvidraco. The highest abundance expression of IGFBP1 transcript was detected in liver, and mRNA level oflGFBP1 in heart, kidney, muscle and intestine from male was higher than that from female （P〈0.05）. The IGFBP1 protein expression from male was higher than that from female. The results provided data support for further functional investigation on 1GFBP1 gene in Pelteobagrusfulvidraco.