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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450bp,编码149个氨基酸残基,推测编码蛋白的分子量为16.9kD,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261bp。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 armigeraPBP2和烟芽夜蛾Heliothis virescensPBP2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。
Pheromone binding proteins (PBPs) play an important role in information exchange between male and female insects. In this study, the open reading frame and the 3’end of the sex pheromone binding protein 2 gene were cloned from the antennae of Helicoverpa assulta (Guenée) by RT-PCR and RACE. The gene was named HassPBP2 (GenBank Accession number is EU316186). The cloning and sequencing results showed that the open reading frame of HassPBP2 was 450 bp in length and encoded 149 amino acid residues. The predicted molecular weight of the protein was 16.9 kD and the isoelectric point was 5.56. HassPBP2 gene structure analysis showed that the gene consists of three exons and two introns, intron length of 90 and 261bp. Amino acid sequence alignment analysis showed that this sequence has the typical characteristic of odor-binding protein, which is in agreement with PBPs of other Lepidoptera insects between 34% and 91%, which is closely related to Helicoverpa armigera PBP2 and Heliothis virescens PBBP2 Sequence consistency up to 91%. Time and tissue expression analysis showed that HassPBP2 was not expressed in the egg stage, larval stage and pupal stage, but expressed in mid-pupa stage and persisted to mid-adult stage and expressed only in the antennae of adult and male adults.