表达序列标签数据库搜索克隆斑马鱼QM基因及其数字化差异显示分析
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浙江大学生命科学学院

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国家自然科学基金,浙江省自然科学基金,高等学校博士学科点专项科研项目


In silico cloning and digital differential display analysis of Danio rerio QM gene by EST database searching
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College of Life Science,Zhejiang University,Hangzhou 310012,China

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    摘要:

    QM基因是一种与肿瘤抑制、细胞生长、分化、发育和凋亡等有关的重要基因,已在人类及许多物种中得到了克隆,但鱼类中尚无报道.我们利用表达序列标签数据库和电子克隆等技术,成功克隆了斑马鱼QM基因,并对该基因的结构进行了系统分析.结果显示,斑马鱼QM基因cDNA全长769 bp,含648 bp开放阅读框架,编码215个氨基酸,5'UTR 25 bp,3'UTR122 bp,所编码的QM多肽分子量24.6 kDa,等电点(pI)10.6,含潜在的蛋白酶C磷酸化位点、N-酰基化位点和酰胺化位点.比较斑马鱼QM和人等13个物种QM的同源性,发现其氨基酸序列相似性为66%~92%.数字化差异显示分析结果表明,QM基因在斑马鱼胚胎及成体多种组织中均有广泛表达.研究结果为今后利用生物信息学快速克隆新的鱼类功能基因打下了方法学基础,也为进一步研究鱼类QM基因功能提供了依据.

    Abstract:

    QM homologs, associated with tumor suppression, cell growth, differentiation, development and apoptosis, were isolated from human and many other organisms. However, QM derived from fish was not reported. Through integrated analysis of public database such as EST, TIGR Gene Index in silico, a full-length eDNA encoding QM homologue of human QM protein was identified from Danio rerio, which is 769 bp length and has a putative protein of 215 amino acids with a predicted mol. wt. of 24.6 kD and isoelectric point of 10.6. It was found that Danio rerio QM amino acid sequence includes 1 Ribosomal protein L10e signature, 2 Amidation sites, 1 Tyrosine sulfation site, 1 N-myristoylation site and 1 Protein kinase C phosphorylation site by scanning PROSITE motifs. The Danio rerio QM shares an 66% - 92% amino acid sequence identical to other proteins of the QM family, including human etc, respectively. Study showed that Danio rerio QM gene expressed in all Danio rerio embryo and adult tissues by digital differential display based on UinGene. The experiment provided us with a feasible bioinformatics method for cloning new functional genes, thereby preparing for further research on fish QM gene function.

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金红建.表达序列标签数据库搜索克隆斑马鱼QM基因及其数字化差异显示分析[J].水产学报,2005,29(5):

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  • 收稿日期:2014-03-09
  • 最后修改日期:2014-03-09
  • 录用日期:2014-03-10
  • 在线发布日期: 2014-03-10
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