›› 2009, Vol. 52 ›› Issue (11): 1199-1205.

• 研究论文 • 上一篇    下一篇

烟夜蛾性信息素结合蛋白2(PBP2)基因的克隆、序列分析与时空表达

李亮, 杨文玲, 郭线茹, 罗梅浩, 原国辉, 乔奇, 付晓伟   

  • 出版日期:2009-12-18 发布日期:2009-11-20

Cloning, sequence analysis and spatio-temporal expression of a pheromone binding protein 2 (PBP2) gene from Helicoverpa assulta (Guenée) (Lepidoptera: Noctuidae)

  • Online:2009-12-18 Published:2009-11-20

摘要: 性信息素结合蛋白(pheromone binding proteins, PBPs)在昆虫雌雄间信息交流中起着重要作用。 本研究利用RT-PCR和RACE方法, 从烟夜蛾Helicoverpa assulta (Guenée)雄虫触角中克隆了性信息素结合蛋白2基因的开放阅读框及3′末端序列, 该基因被命名为HassPBP2(GenBank登录号为EU316186)。克隆和测序结果表明, HassPBP2开放阅读框全长450 bp, 编码149个氨基酸残基, 推测编码蛋白的分子量为16.9 kD, 等电点为5.56。HassPBP2基因结构分析表明, 该基因由3个外显子和2个内含子组成, 内含子的长度分别为90和261 bp。氨基酸序列联配分析表明, 此序列具有气味结合蛋白的典型特征, 与其他鳞翅目昆虫PBPs的一致性在34%~91%之间, 其中与棉铃虫Helicoverpa armigera PBP2和烟芽夜蛾Heliothis virescens PBP2的序列一致性高达91%。时间表达和组织表达分析显示, HassPBP2在卵期、幼虫期和蛹早期不表达, 在蛹中期开始表达, 并一直持续到成虫中期, 且只在雌、雄成虫触角中表达。

关键词: 烟夜蛾, 性信息素结合蛋白, 基因克隆, 序列分析, 时空表达

Abstract: Pheromone binding proteins (PBPs) play an important role in communication between male and female insects. The open reading frame and 3′ end full-length cDNA of a novel pheromone binding protein 2 gene was cloned from Helicoverpa assulta (Guenée) by using RT-PCR and RACE methods, which was named HassPBP2 (GenBank accession no. EU316186). The cloning and sequencing results showed that the full-length of open reading frame in HassPBP2 was 450 bp, encoding 149 amino acid residues with the predicted molecular weight and isoelectric point of 16.9 kD and 5.56, respectively. Gene structure analysis revealed that HassPBP2 consists of 3 exons and 2 introns with length of 90 and 261 bp, respectively. Amino acid sequence analysis showed that the deduced amino acid sequence of HassPBP2 had the characteristics of odorant binding proteins. HassPBP2 had 34%-91% identity with PBPs of other Lepidoptera moths, and shared 91% identity with the PBP2 sequences from H. armigera and Heliothis virescens. Temporal expression revealed that HassPBP2 transcript was observed clearly from mid-stage pupa to mid-stage adult, but not in the egg, larva and early-stage pupa. The tissue analysis further indicated that HassPBP2 was only expressed in the antennae of both male and female adults.

Key words: Helicoverpa assulta, pheromone binding protein, gene cloning, sequence analysis, spatio-temporal expression