团头鲂SREBP1基因的克隆及其表达分析
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国家自然科学基金项目(31401976)


Cloning and expression of SREBP1 gene in Megalobrama amblycephala
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    摘要:

    为阐明团头鲂SREBP1的功能,采用RT-PCR技术克隆获得SREBP1基因的cDNA序列,利用荧光定量 PCR(qRT-PCR)检测SREBP1基因在不同发育时期及不同组织中的表达情况。生物信息学分析发现,SREBP1基因cDNA的编码序列(coding sequence,CDS)为3 426 bp(GenBank登录号:MH633449),开放阅读框(ORF)编码1 141个氨基酸,与大西洋鲑、斑马鱼、草鱼、鲤、金线鲃SREBP1的氨基酸序列相似性较高(74%~99%)。保守结构域分析显示,团头鲂SREBP1蛋白具有SREBPs家族典型的“碱性螺旋环螺旋亮氨酸拉链”(bHLHZip)结构域。表达分析结果显示,SREBP1基因在团头鲂胚胎发育过程中的表达呈现先下降后增高的趋势,在眼囊期最低,至出膜期达到最高。在团头鲂幼鱼中,SREBP1在脑组织中的表达量最高,其次是眼、肝脏、心脏、肌肉、脂肪组织、肾脏,鳃中表达量最低。在成鱼中,SREBP1在脑组织表达量最高,其次是性腺组织、肌肉、肝脏、眼,肾脏、脾脏和心脏中的表达量较低。以上结果表明,SREBP1基因在团头鲂幼鱼和成鱼的不同组织中的表达模式存在差异。

    Abstract:

    This study was conducted to investigate the function of the SREBP1 gene inMegalobrama amblycephala. The SREBP1 gene was cloned by RTPCR,and its expression were analyzed using qRTPCR. Bioinformatics analysis showed that the cDNA coding sequence (CDS) of SREBP1 was 3 426 bp (GenBank Accession number:MH633449),with an open reading frame (ORF) encoding 1 141 amino acids. The M. amblycephala SREBP1 protein shared high similarity (74%~99%) with Salmo salar,Danio rerio,Ctenopharyngodon idella,Cyprinus carpio and Sinocyclocheilus grahami. Conserved domain analysis showed that the SREBP1 protein has the bHLHZip domains,which belong to the SREBPs family. Expression analysis showed that the SREBP1 expression leveled was the lowest at the optic vesicle stage and highest at the hatching stage during embryonic development of M. amblycephala. For juvenileM. amblycephala,the SREBP1 gene expressed highest in the brain,followed by the eye,liver,heart,fat tissue,kidney and gill. For adultM. amblycephala,theSREBP1 gene expressed highest in the brain,followed by the gonad,muscle,liver,eye,while expressed relatively low in the kidney,spleen and heart. The tissue expression pattern of the SREBP1 gene was different between the juvenile and adult M. amblycephala.

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吴文新,曹赞,王凡,徐佳,王卫民,赵玉华.团头鲂SREBP1基因的克隆及其表达分析[J].华中农业大学学报,2019,38(2):9-16

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  • 收稿日期:2018-07-20
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  • 在线发布日期: 2019-01-30
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