桔小实蝇非典型气味受体 Orco 基因的克隆与表达谱分析
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国家自然科学基金项目(31071713)和教育部博士学科点科研基金项目(20094404110019)


Gene cloning and expression profiling of atypical odorant receptors Orco gene in Bactrocera dorsalis (Hendel)
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    摘要:

    为探讨非典型气味受体基因 Orco 的功能,采用RT-PCR 和RACE 方法克隆获得桔小实蝇 Bactocera dorsalis (Hendel) Orco 受体的全长cDNA 序列,命名为 Bdor/Orco (GenBank登录号为EU621792)。测序结果表明 Bdor/Orco 开放阅读框全长1 422 bp,编码473个氨基酸残基。对其组成成分、疏水/亲水区域、信号肽、蛋白质二级结构,以及分子进化关系等进行了预测与推断,分析结果表明 Bdor/Orco 与其他昆虫的 Orco 具有较高的氨基酸序列一致性,特别是C末端。对该基因在桔小实蝇成虫不同组织和发育时期表达量的荧光定量PCR分析,结果表明 Bdor/Orco 主要是在桔小实蝇成虫触角中表达,头部(去除触角)、雌虫前足和翅中也有较高的表达; 桔小实蝇在各个发育时期的表达水平不同,在刚羽化雌成虫中的表达量最高。

    Abstract:

    In order to explore the functions of Orco , the gene encoding Orco from Bactrocera dorsalis (Hendel) was cloned and named as Bdor/Orco (GenBank accession No.EU621792) by RT-PCR and RACE. The sequencing results showed that the open reading frame of Bdor/Orco was 1 422 bp in length,encoding 473 amino acid residues. The Bdor/Orco was analyzed and predicted by the tools of bioinformatics in the following aspects: the composition, hydrophobicity or hydrophilicity, the signal peptide, protein secondary structure, the composition of homologous protein and the phylogenetic relationship. The results showed Bdor/Orco had the characteristics of Orco , with special conserved C domains. Tissue expression pattern revealed that Bdor/Orco transcript was observed clearly in the antenna of B.dorsalis , and highly expressed in the head (with antennae removed), female foreleg and wing. The temporal expression pattern further indicated that Bdor/Orco was also expressed in different developmental stages at different expression levels with the highest level in newly emerged female adult.

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赵海明,易欣,胡美英,安国栋,王会冬,黄林.桔小实蝇非典型气味受体 Orco 基因的克隆与表达谱分析[J].华中农业大学学报,2013,32(1):40-47

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