中华按蚊气味结合蛋白基因AsinOBP1的克隆和表达分析

摘要/Abstract

摘要： 【目的】蚊虫的行为在很大程度上依赖于嗅觉系统, 例如寻找宿主和产卵场所等。中华按蚊Anopheles sinensis是我国最重要的传疟媒介之一, 但有关中华按蚊嗅觉信号传递过程的研究甚少。本研究旨在克隆和表达分析中华按蚊的气味结合蛋白（odorant binding proteins, OBPs）基因, 为进一步研究中华按蚊嗅觉传递的分子机制奠定基础。【方法】通过分析中华按蚊的转录组数据克隆气味结合蛋白基因, 采用RT-PCR和实时定量PCR技术分析该基因在成虫不同组织和在吸血前后的表达模式。【结果】克隆到一个气味结合蛋白基因, 命名为AsinOBP1 (GenBank登录号为KJ958382)。AsinOBP1基因开放阅读框长435 bp, 编码144个氨基酸, 具有典型的6个半胱氨酸位点。RT-PCR组织表达谱分析发现, AsinOBP1在检测的所有成虫触角、下颚须、喙和头部组织中都有表达, 而在足和去掉头部以外的躯体组织中不表达。定量分析发现AsinOBP1在雌蚊触角中的表达水平最高, 吸食血液后, AsinOBP1的表达水平显著下降；仅用小鼠气味处理后, AsinOBP1的表达水平也显著下降。【结论】研究结果说明AsinOBP1可能是嗅觉组织特异性表达的基因, 与雌蚊寻找宿主等行为有关, 其功能还需深入研究。

关键词: 中华按蚊, 气味结合蛋白, 组织表达谱, 定量PCR, 吸血

Abstract: 【Aim】 Olfactory cues play a critical role in mediating a variety of important behaviors in mosquitoes including host seeking, and oviposition. Anopheles sinensis is the major malaria vector in China. However, little is known about the olfactory signal transmission of An. sinensis. Our study aims to clone an odorant binding protein (OBP) gene from An. sinensis and analyze its expression patterns in different tissues and in response to blood-meal feeding, so as to provide the foundation for further research of the molecular mechanism of An. sinensis olfactory transmission. 【Methods】 A transcriptome database for An. sinensis was mined through bioinformatic analysis. Gene expression levels were analyzed in different tissues of An. sinensis adults and at different time post blood-meal feeding using reverse transcription PCR (RT-PCR) or real-time PCR. 【Results】 An odorant binding protein gene was identified and named AsinOBP1, with the Genbank accession number of KJ958382. AsinOBP1 contains a 453-bp open reading frame encoding 144-amino-acid residues including an N-terminal signal peptide. The mature protein of AsinOBP1 contains six conserved cysteine residues, which are the hallmark of insect OBPs. Tissue expression profile analysis showed that AsinOBP1 was expressed in all tested tissues of antennae, maxillary palps, proboscis and head in adults, but not in tissues of legs and body (head removed). The expression level of AsinOBP1 in antennae of female adults was strongly downregulated after blood-meal feeding. Similarly, AsinOBP1 expression in antennae of female adults was also decreased significantly in response to mouse odor. 【Conclusion】 These findings strongly suggest that AsinOBP1 might be expressed specifically in olfactory tissues and involved in host seeking. Further functional analysis is required to clarify the roles of AsinOBP1 in An. sinensis.

Key words: Anopheles sinensis, odorant binding protein, tissue expression pattern, quantitative real-time PCR, blood-meal feeding

秦赠, 冉永红, 支中婧, 闫振天, 张玉娟, 黄婷, 何正波, 陈斌. 中华按蚊气味结合蛋白基因AsinOBP1的克隆和表达分析[J]. , 2014, 57(11): 1289-1298.

QIN Zeng, RAN Yong-Hong, ZHI Zhong-Jing, YAN Zhen-Tian, ZHANG Yu-Juan, HUANG Ting, HE Zheng-Bo, CHEN Bin. Cloning and expression analysis of an odorant binding protein gene AsinOBP1 from Anopheles sinensis (Diptera: Culicidae)[J]. , 2014, 57(11): 1289-1298.

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中华按蚊气味结合蛋白基因AsinOBP1的克隆和表达分析

Cloning and expression analysis of an odorant binding protein gene AsinOBP1 from Anopheles sinensis (Diptera: Culicidae)

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