光肩星天牛性信息素结合蛋白AglaPBP1和AglaPBP2基因鉴定和表达分析
Identification and expression patterns of the pheromone binding protein genes AglaPBP1 and AglaPBP2 in Anoplophora glabripennis (Coleoptera: Cerambycidae)
王菁桢** 胡 平 骆有庆 陶 静***
点击：1836次 下载：8次
DOI：10.7679/j.issn.2095-1353.2017.005
作者单位：北京林业大学林木有害生物防治北京市重点实验室，北京 100083
中文关键词：光肩星天牛，性信息素结合蛋白，克隆，表达谱分析
英文关键词：Anoplophora glabripennis, pheromone binding proteins, cloning, expression analysis
中文摘要：
【目的】 为了鉴定光肩星天牛Anoplophora glabripennis (Motsch.)的性信息素结合蛋白PBPs基因并明确其在雌雄成虫不同部位的表达差异。【方法】 本文基于光肩星天牛触角转录组测序数据，通过blast比对得到2条PBPs基因片段，克隆测序得到2条PBPs基因的全序列，并进行生物信息学分析。采用Real-time PCR 方法分析了2条PBPs基因在光肩星天牛雌雄成虫间的表达差异。【结果】 克隆得到性信息素结合蛋白基因，命名为AglaPBP1（GenBank登录号：KX272639）和AglaPBP2（GenBank登录号：KX272640）。AglaPBP1开放阅读框长为411 bp，编码136个氨基酸，预测分子量为15.02 ku，等电点为4.22，AglaPBP2开放阅读框长为408 bp，编码135个氨基酸，预测分子量为15.00 ku，等电点为5.16。AglaPBP1和AglaPBP2都有完整的信号肽，分别位于1-21位氨基酸和1-19位氨基酸。AglaPBP1和 AglaPBP2氨基酸序列中均有6个保守的半胱氨酸残基，与云斑天牛Batocera horsfield i(Hope) BhorPBP1和BhorPBP2的同源性分别为74%和49%。qPCR结果显示：AglaPBP1在雌雄虫触角、下颚须，以及雌虫足中均有的表达，且雌虫触角表达量高于雄虫。AglaPBP2仅在雄虫触角中显著表达，在雌雄虫的下颚须中也有少量表达。【结论】 克隆获得了AglaPBP1和AglaPBP2基因序列，并明确了这两个基因在成虫不同部位的表达情况，为进一步研究其功能，从而为更好地了解PBPs在光肩星天牛嗅觉识别过程中的作用奠定了基础。
英文摘要：
[Objectives] To identify pheromone binding protein (PBP) genes in Anoplophora glabripennis (Motsch.) and detect their differential expression in different organs in males and females of this species. [Methods] Two PBP gene fragments were obtained by blast based on a previously established antennal transcriptome. Complete sequences were obtained by cloning and sequencing, after which bioinformatic methods were employed. Real-time PCR was used to detect the expression patterns of PBP genes in different sexes. [Results] Two PBP genes were obtained from A. glabripennis; AglaPBP1 (GenBank accession no. KX272639) and AglaPBP2 (GenBank accession no.KX272640). The open reading frame of AglaPBP1 is 411 bp, encoding 136 amino acid residues with a predicted molecular weight and isoelectric point of 15.02 ku and 4.22, respectively. The open reading frame of AglaPBP2 is 408 bp, encoding 135 amino acid residues with a predicted molecular weight and isoelectric point of 15.007 ku and 5.16, respectively. AglaPBP1 and AglaPBP2 both have signal peptides at 1-21 and 1-19. Both AglaPBP1 and AglaPBP2 are characterized by six conservative cysteine (Cys) residues. The two genes have about 74% and 49% similarity with the PBP genes of Batocera horsfieldi (Hope). qPCR analysis indicated that AglaPBP1 was expressed in the antennae, maxillary palps and legs, and that expression in female antennae was higher than in male antennae. AglaPBP2 was almost specifically expressed in male antennae. [Conclusion] Clarifying AglaPBP1 and AglaPBP2 expression in different organs lays a foundation for further study of the function of these genes and the molecular mechanisms underlying insect olfaction in general.