胆固醇损伤内皮细胞差异表达基因的生物信息学分析
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国家自然科学基金(3002103);湖南省自然科学基金(02JJY4011);湖南省教育厅青年基金(02B039)


Bioinformatic Analysis of Differentially Displayed Gene in Human Endothelial Cell Induced by Cholesterol
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    摘要:

    目的利用差异表达基因克隆方法(抑制消减杂交)获得大量的动脉粥样硬化相关候选基因和表达序列标签后,探讨如何进行后续基因表达及功能的研究。方法利用Internet网络上的数据库及生物学分析软件对胆固醇损伤内皮细胞后获得的差异表达基因进行核酸序列和蛋白质序列分析,探索差异表达基因克隆后的研究方法和思路。结果通过电子延伸得到一个684bp的全长cDNA序列;通过核酸序列分析,该序列定位在线粒体基因组的7587位~8270位,含有一个完整的开放阅读框,编码与氧化磷酸化相关的9个亚基。对其中一个亚基细胞色素氧化酶Ⅱ(COX2)分析得知,细胞色素氧化酶Ⅱ基因编码一段25.6kDa的弱酸性的信号锚蛋白,细胞色素氧化酶Ⅱ蛋白的三维结构是一个典型的椅式结构,它含有一段疏水区域,一个跨膜结构域和一个胞质结构域。运用蛋白的进化分析,得知细胞色素氧化酶Ⅱ蛋白胞质结构域的氨基酸序列在进化过程中高度保守。结论生物信息学技术是一种高效的获取疾病相关基因信息的方法,利用生物信息学方法对细胞色素氧化酶Ⅱ基因进行分析,获得了基因及其编码蛋白的相关信息,该蛋白参与电子传递,可能与细胞的氧化应激有关。

    Abstract:

    Aim To study the large amount of differentially displayed genes and expressed sequence-tags (EST) obtained by gene clone is a difficult problem. It is important to exploit new methods for further study. Methods EST were acquired from human umbilical vein endothelial cells induced by cholesterol by suppression subtractive hybridization (SSH). Bioinformatics analysis of atherosclerosis related genes was done by the bioinformatical databases and bio-softwares such as BLAST, ExPasy analyse soft box, DNAMAN soft, BioEdit soft and RasMol soft. Results We obtained a 684 bp complete cDNA sequence by electronic clone, which was homogeneous to cytochrome C oxidase subunit Ⅱ gene (COX2). The complete genome of COX2 sited in homo sapiens mitochondrion, and included a complete open reading frame (ORF) coding 227 amino acids. The analysis of its protein sequence indicated that COX2 gene encode a 25.6 kDa protein, which was a weak acid signal anchor. The three-dimensional structure of COX2 was a representative chair-like stucture, containing a hydrophobicity region, a transmembrane domain and a periplasmic domain. The periplasmic domain of the COX2 protein sequence was high conservative in the process of evolutions by the analysis of the protein evolutions. Conclusion We obtained the information of COX2 about nucleic acid sequence and protein sequence by bioinformatics analysis

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彭瑾瑜,朱勋,唐蔚青,王抒,黎健,王仁,郭芳,刘俊文,杨向东.胆固醇损伤内皮细胞差异表达基因的生物信息学分析[J].中国动脉硬化杂志,2005,13(3):259~262.

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  • 收稿日期:2004-06-24
  • 最后修改日期:2005-02-28
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