悬铃木方翅网蝽非典型气味受体基因CcilOrco的克隆与序列分析  

Gene cloning and sequence analysis of the odorant receptor gene CcilOrco in Corythucha ciliata

作　　者：董敏 杜立啸 袁洪振 夏延颖 司军红 董文慧 DONG Min;DU Lixiao;YUAN Hongzhen;XIA Yanying;SI Junhong;DONG Wenhui(Shandong Honglin Engineering Technology Co.,Ltd.,Jinan 250101,China 2.Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China)

机构地区：[1]山东鸿林工程技术有限公司,济南250101 [2]中国农业科学院植物保护研究所,北京100193

出　　处：《植物保护》2018年第4期53-59,共7页Plant Protection

摘　　要：为进一步研究悬铃木方翅网蝽Corythucha ciliate嗅觉通讯分子机制和寻求新的悬铃木方翅网蝽防治技术,本研究克隆了悬铃木方翅网蝽的非典型气味受体基因CcilOrco,并对其序列进行生物信息学分析。根据GenBank中已发表的半翅目昆虫非典型气味受体家族基因的氨基酸保守序列设计简并引物,采用RT-PCR方法扩增目的基因,将其克隆至pEASY-Blunt载体并测序。将克隆获得悬铃木方翅网蝽非典型气味受体Orco的cDNA序列命名为CcilOrco(GenBank登录号:MF564288),序列分析结果显示,CcilOrco开放阅读框长1 419bp,编码472个氨基酸。预测其分子量为53.25kD,等电点为6.22,序列中有7个跨膜区,N-端在细胞膜内,C-端在细胞膜外。通过在GenBank中进行序列的同源性比较,该基因与已公布的半翅目昆虫的非典型气味受体基因序列有较高的同源性。克隆所获得的基因属于非典型气味受体家族基因。qPCR结果显示CcilOrco主要在雌雄成虫触角中高表达。For further studying the mechanism of olfactory communication and seeking new prevention technology for Corythucha ciliate,the olfactory receptor gene（CcilOrco）was cloned and analyzed by bioinformatics analysis.According to the conserved amino acid sequences of the Hemiptera insect olfactory receptor gene family published in GenBank,the degenerate primers were designed for RT-PCR.The cDNA sequence of Corythucha ciliate olfactory receptor gene was cloned,and named CcilOrco（GenBank accession no：MF564288）.The results of sequence analysis showed that the open reading frame of CcilOrco consisted of 1 419 nucleotides,encoding 472 amino acids with a molecular weight of 53.25 kD,an isoelectric point of 6.22 and7 putative transmembrane domains in the sequence,with an intracellular N-terminal and extracellular C-terminus.By comparing the homology of the sequences in GenBank,the sequence shared high identity with Orco orthologue chemoreceptor sequences from previously reported insects.The results indicated that the gene belonged to atypical olfactory receptor gene family.qPCR showed that CcilOrco was expressed highly in male and female antennae.

关 键 词：悬铃木方翅网蝽 气味受体 基因克隆 序列分析 表达分析 

分 类 号：S763.7[农业科学—森林保护学]