缢蛏IGFBP基因结构及生长性状相关SNP筛选
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上海海洋大学,上海海洋大学,上海海洋大学,上海海洋大学,上海海洋大学,上海海洋大学,上海海洋大学

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国家“八六三”高技术研究发展计划(2012AA10A400);国家自然科学基金(31472278);上海知识服务平台(ZF1206)


Molecular characterization of IGFBP and association analysis with growth traits in the razor clam Sinonovacula constricta
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Key Laboratory of Freshwater Germplasm Resources,Ministry of Agriculture,Shanghai Ocean University;,,,,,

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    摘要:

    类胰岛素生长因子结合蛋白(IGFBP)是IGF系统的一部分,主要参与IGF的运输、定位和生物活性调节。本研究采用RACE技术和长PCR技术,克隆了缢蛏IGFBP基因的cDNA和DNA全长序列,应用荧光定量PCR技术分析了缢蛏不同发育时期和不同组织中IGFBP mRNA的表达特征,并进一步筛选了IGFBP基因与生长性状相关的SNP位点。序列分析表明,缢蛏IGFBP cDNA序列全长631 bp,包括5'端非编码区60 bp,3'端非编码区136 bp和开放阅读框435 bp,编码144个氨基酸。该基因含有保守的IGFBP-N端,包含12个半胱氨酸残基,其中1~18个氨基酸为信号肽,属于分泌型蛋白。IGFBP DNA全长3 122 bp,其中包含1个内含子(2 687 bp)和2个外显子(200和235 bp)。荧光定量PCR结果显示,IGFBP mRNA在消化腺组织中表达量最高;在缢蛏的稚贝期,IGFBP mRNA呈现高表达,而在其他发育时期表达量低。在IGFBP基因中筛选到4个SNP位点,其中1个SNP位点与缢蛏的壳长和体质量呈显著相关。

    Abstract:

    Insulin like growth factor binding protein(IGFBP)is a member of IGFBP family participating in transportation,localization and biological regulation of IGFs.Here,we conducted RACE-PCR and LONG-PCR for the cloning of full length cDNA and DNA of IGFBP gene in Sinonovacula constricta.Quantitative real-time PCR analysis revealed that IGFBP transcript was expressed in a wide range of tissues and developmental stages.Additional growth-linked SNP were discovered based on association study.The full length of IGFBP cDNA sequence consists of 631 base pairs(bp),comprising a 60 bp 5' untranslated region(UTR),a 136-bp 3' UTR,and an open reading frame(ORF)which is 435 bp in length.The ORF encodes 144 amino acids with a signal peptide(18 amino acids)and 12 cysteine residues in the conserve N-terminal.There is one intron(2 687 bp)and two exons(200 and 245 bp)in the sequence of IGFBP gene(3 122 bp).Based on quantitative real-time PCR analysis,we found the highest level of IGFBP expression in the digestive gland.Also the transcript was highly detected in juvenile stages rather than other stages.Four SNPs were identified in IGFBP genomic DNA and one of them was significantly associated with the shell length and body weight.Our results indicated that IGFBP gene plays important roles in the growth and development in S.constricta.

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谢淑媚,牛东红,阮海灯,王泽,王飞,陈世娥,李家乐.缢蛏IGFBP基因结构及生长性状相关SNP筛选[J].水产学报,2015,39(6):799~809

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  • 收稿日期:2014-12-24
  • 最后修改日期:2015-03-13
  • 录用日期:2015-06-15
  • 在线发布日期: 2015-06-23
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