中国农业科学 ›› 2010, Vol. 43 ›› Issue (3): 618-625 .doi: 10.3864/j.issn.0578-1752.2010.03.024

• 研究简报 • 上一篇    下一篇

2个新的鳞翅目OR83b类化感蛋白基因的克隆和序列分析

修伟明,张逸凡,杨殿林,董双林,刘玉升

  

  1. (农业部环境保护科研监测所/转基因生物生态环境安全监督检验测试中心)
  • 收稿日期:2009-05-04 修回日期:2009-08-11 出版日期:2010-02-10 发布日期:2010-02-10
  • 通讯作者: 董双林

Molecular Cloning and Sequence Analysis of Two cDNA Genes of Two New Lepidoptera OR83b Orthologue Chemoreceptors

XIU Wei-ming, ZHANG Yi-fan, YANG Dian-lin, DONG Shuang-lin, LIU Yu-sheng
  

  1. (农业部环境保护科研监测所/转基因生物生态环境安全监督检验测试中心)
  • Received:2009-05-04 Revised:2009-08-11 Online:2010-02-10 Published:2010-02-10
  • Contact: DONG Shuang-lin

摘要:

【目的】分离和克隆昆虫触角中一类非常保守的嗅觉受体蛋白基因全序列。【方法】采用简并引物反转录多聚酶链式反应(RT-PCR)和cDNA末端快速扩增(RACE)技术克隆基因,用生物信息学方法对获得的cDNA全序列及推导的氨基酸序列进行分析。【结果】从甜菜夜蛾(Spodoptera exigua Hübner)和斜纹夜蛾[Spodoptera litura (Fabricius)]雄蛾触角中扩增得到2个分别为1 906 bp和2 483 bp的OR83类化感蛋白基因的cDNA,开放阅读框(ORF)编码473个氨基酸,命名为SexiOR2和SlitOR2。通过在GenBank中进行序列的同源性比较,发现这2个鳞翅目化感蛋白新序列与已知蛾类昆虫的OR83b类化感蛋白氨基酸序列具较高的同源性。【结论】明确了所获得的2个鳞翅目蛋白新序列属于OR83b类化感蛋白。

关键词: 甜菜夜蛾, 斜纹夜蛾, 嗅觉受体基因, 序列分析

Abstract:

【Objective】 The aim of this study was to isolate the cDNA full sequence of an unusually highly conserved olfactory receptor (orthologue to the Drosophila melanogaster DOR83b) gene from the antennae of insects. 【Method】 The olfactory receptor genes were cloned by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. Bioinformatics methods were used to further analyze cDNA sequence obtained and putative amino acid sequence. 【Result】 Two cDNA full sequences of olfactory receptor (OR) gene were amplified from the antennae of male Spodoptera exigua and S. litura, and they were named SexiOR2 and SlitOR2. SexiOR2 and SlitOR2 contained 1 906 bp and 2 483 bp, and with deduced amino acid sequences of 473 residues. The sequence analysis indicated that the deduced amino acid sequences of the cDNA sequences shared high identity with OR83b orthologue chemoreceptor sequences from other previously reported moths. 【Conclusion】 It is inferred that the sequences obtained are cDNA sequences of OR83b orthologue chemoreceptor gene.

Key words: Spodoptera exigua, Spodoptera litura, olfactory receptor gene, sequence analysis