牙鲆胰岛素样生长因子结合蛋白IGFBP-1 cDNA全长的克隆及表达分析
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上海海洋大学农业部水产种质资源和养殖生态重点开放实验室,上海海洋大学农业部水产种质资源和养殖生态重点开放实验室,上海海洋大学农业部水产种质资源和养殖生态重点开放实验室

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国家自然科学(30571420);上海市重点学科水生生物学建设项目(S30701)资助


The full-length cDNA cloning and gene expression analysis of insulin-like growth factor binding protein 1(IGFBP-1)in Japanese flounder(Paralichthys olivaceus)
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Key Laboratory of Aquatic Genetic Resources and Utilization Ministry of Agriculture Shanghai Ocean University,Key Laboratory of Aquatic Genetic Resources and Utilization Ministry of Agriculture Shanghai Ocean University,Key Laboratory of Aquatic Genetic Resources and Utilization Ministry of Agriculture Shanghai Ocean University

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    摘要:

    利用RACE-PCR技术,从牙鲆(Paralichthys olivaceus)的肝脏组织总RNA中克隆得到胰岛素样生长因子结合蛋白-1(insulin-like growth factor binding protein 1, IGFBP-1)基因的全长cDNA序列,该cDNA全长为1070 bp,开放阅读框为729 bp,编码242个氨基酸。通过系统进化树分析,牙鲆IGFBP-1与鱼类IGFBP-1基因聚为一支;通过同源性比对,牙鲆IGFBP-1基因的核苷酸序列与大菱鲆同源性最高,为95%,而其推导的氨基酸序列与其它鱼类如大菱鲆、五条鰤、黄金鲈、红点鲑、鲤鱼和斑马鱼的同源性分别为89%、89%、84%、79%、67%和67%。半定量RT-PCR分析表明,牙鲆IGFBP-1基因存在母源转录本,合子基因在孵化前的胚胎阶段及早期仔鱼中仅有较低水平的表达,在后期仔鱼中表达逐渐增高;牙鲆IGFBP-1基因在肝脏中表达量最高,在胃、脾、肠、性腺、肾、鳃、脑、心脏和肌肉中也有不同程度的表达。

    Abstract:

    The full-length cDNA of insulin-like growth factor binding protein 1 gene(IGFBP-1) was obtained from the liver of Japanese flounder(Paralichthys olivaceus)using RACE method. The complete cDNA of P.olivaceus IGFBP-1 is 1 070 bp and its ORF includes 729 bp encoding 242 amino acid residues.The IGFBP-1 was highly homologous with IGFBP-1 gene in fish through phylogency analysis. The nucleotide sequence of IGFBP-1 from P.olivaceusshared 95% homology with that of Scophthalmus maximu,and the deduced amino acid analysis showed that the sequence similarities betweenP.olivaceusand S.maximu,Seriola quinqueradiata,Perca flavescens,Salvelinus alpines,Cyprinus carpio andDanio rerio,are 89%,89%,84%,79%,67% and 67%,respectively. Semi-quantitative RT-PCR results demonstrated that IGFBP-1 mRNA was detected in unfertilized egg,fertilized egg,embryonic and larval development,and various adult tissues.Moreover,real-time quantitative RT-PCR results showed that IGFBP-1 gene had the highest expression in adult liver and was expressed in a stage-specific manner during embryonic and larval development.

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翟万营,张俊玲,施志仪,李巍.牙鲆胰岛素样生长因子结合蛋白IGFBP-1 cDNA全长的克隆及表达分析[J].水产学报,2012,36(2):170~179

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  • 收稿日期:2011-07-16
  • 最后修改日期:2011-11-07
  • 录用日期:2012-01-14
  • 在线发布日期: 2012-02-16
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