›› 2015, Vol. 58 ›› Issue (6): 603-609.

• 研究论文 • 上一篇    下一篇

棉铃虫齿唇姬蜂嗅觉受体基因 CchlOrco 的克隆及组织表达谱分析

董钧锋1, 宋月芹1, 孙九光2, 闫志宁1, 孙亚兰3, 王琛柱3,*   

  1. (1. 河南科技大学林学院, 河南洛阳 471003; 2. 安阳工学院生物与食品工程学院, 河南安阳 455000; 3. 中国科学院动物研究所, 农业虫害鼠害综合治理研究国家重点实验室, 北京 100101)
  • 出版日期:2015-06-20 发布日期:2015-06-20
  • 作者简介:董钧锋, 男, 1973年生, 河南许昌人, 博士, 副教授, 研究方向为农业昆虫与害虫防治, E-mail: junfengdong@haust.edu.cn

Cloning and tissue expression profiling of the olfactory receptor gene CchlOrco in Campoletis chlorideae (Hymenoptera: Ichneumonidae)

DONG Jun-Feng1, SONG Yue-Qin1, SUN Jiu-Guang2, YAN Zhi-Ning1, SUN Ya-Lan3, WANG Chen-Zhu3,*   

  1. (1. Forestry College, Henan University of Science and Technology, Luoyang, Henan 471003, China; 2. Biology and Food Engineering College, Anyang Institute of Technology, Anyang, Henan 455000, China; 3. State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China)
  • Online:2015-06-20 Published:2015-06-20

摘要: 【目的】昆虫的嗅觉受体(olfactory receptors, ORs)一般以气味分子特异的ORs与共受体( co-Receptor, Orco)通过形成异质二聚体在嗅觉感受中发挥关键作用,其中Orco由于具有序列的保守性而受到广泛的重视。本研究旨在克隆棉铃虫齿唇姬蜂 Campoletis chlorideae 的Orco基因,并对其组织表达谱进行分析。【方法】利用RT-PCR技术和转录组分析技术克隆棉铃虫齿唇姬蜂的Orco基因,并对其编码的氨基酸序列进行生物信息学分析;利用Real-time PCR技术对该基因在该蜂成虫不同组织中的表达量进行分析。【结果】获得了棉铃虫齿唇姬蜂 Orco 的全长cDNA序列,命名为 CchlOrco(GenBank登录号:KP255444)。序列分析表明, 该基因开放阅读框全长1 437 bp,编码478个氨基酸,预测该氨基酸序列具有7个跨膜区。CchlOrco 主要在成虫触角中表达,且在雄蜂触角中的表达量最高,是雌蜂触角中表达量的8.0倍,而在其他组织中表达量极低。【结论】本研究克隆了棉铃虫齿唇姬蜂 CchlOrco 序列全长,明确了其在成虫不同组织中的表达水平,为进一步研究该基因及其他嗅觉受体基因功能奠定了基础。

关键词: 棉铃虫齿唇姬蜂, 嗅觉受体, 基因克隆, 组织特异性, 荧光定量PCR

Abstract: 【Aim】 Olfactory receptors (ORs) interact with the co-receptor (Orco) to form heterodimeric complexes that are critical for insect olfaction. Due to its highly conserved sequence, Orco has received extensive attention. The present study aims to clone and identify the Orco gene in Campoletis chlorideae, and analyze its tissue expression profiles. 【Methods】 The Orco gene from C. chlorideae was cloned by RT-PCR and transcriptome technology, and the putative amino acid sequence was analyzed by bioinformatics methods. The expression level of Orco in different tissues of C. chlorideae adults was analyzed by realtime quantitative PCR. 【Results】 The full-length cDNA sequence of Orco was obtained from C. chlorideae, and named as CchlOrco (GenBank accession no. KP255444). Sequence analysis revealed that the open reading frame of CchlOrco is 1 437 bp in length, encoding 478 amino acid residues with seven transmembrane domains. Real-time quantitative PCR results indicated that CchlOrco was mainly expressed in the adult antennae of C. chlorideae, while expressed in other tissues at an extremely low level. The expression level of CchlOrco in the antennae of male adults was 8.0-fold as high as that in the antennae of female adults. 【Conclusion】 The cloning of the full-length olfactory receptor gene CchlOrco and its expression profiles provide a basis for future functional study of CchlOrco and other olfactory receptor genes of C. chlorideae.

Key words: Campoletis chlorideae, olfactory receptor, gene cloning, tissue-specific expression, real-time quantitative PCR