长足大竹象信息素结合蛋白CbuqPBP1克隆和表达分析  

Molecular Cloning and Expression Analysis of CbuqPBP1 Gene in the Bamboo Snout Beetle,Cyrtotrachelus buqueti

作　　者：杨桦[1] 苏婷[1] 杨伟[1] 杨春平[1] 周学莉 李一平 

机构地区：[1]四川农业大学林学院/四川省林业生态工程省级重点实验室,成都611130 [2]绵阳市林业局,四川绵阳621000

出　　处：《四川农业大学学报》2018年第1期78-85,共8页Journal of Sichuan Agricultural University

基　　金：国家自然科学基金项目(31400548);中国博士后科学基金面上资助项目(2014M560725)

摘　　要：【目的】克隆和表达长足大竹象Cyrtotrachelus buqueti Guerin-Meneville信息素结合蛋白基因并进行基因序列和表达分析,为进一步研究该基因的生理功能奠定基础。【方法】通过RT-PCR技术克隆了信息素结合蛋白基因,采用生物信息方法分析其序列特征,利用实时荧光定量PCR技术研究其在长足大竹象不同虫态和不同组织中的表达量。【结果】克隆获得长足大竹象信息素结合蛋白基因,命名为Cbuq PBP1(Gen Bank登录号:KU845733.1),开放阅读框为432bp,编码143个氨基酸残基,分子量为15.84 k D,等电点为4.60,含6个保守半胱氨酸位点。系统进化树结果显示,Cbuq PBP1与其他昆虫PBP具有较高的相似性。实时荧光定量PCR结果表明,Cbuq PBP1在雄虫、雌虫、幼虫体内均有表达,但在雄虫体内表达量最高(P<0.05)。同时,在雄虫触角中的表达量显著高于其他组织(P<0.05)。【结论】表明Cbuq PBP1在长足大竹象不同虫态和不同组织中差异表达。【Objective】This study aimed to clone and express pheromone binding proteins(PBPs) of Cyrtotrachelus buqueti Guerin-Meneville and to conduct gene sequence and expression analysis,which may lay a solid foundation for further researches of its physical functions.【Method】RT-PCR technology was used to clone the pheromone binding protein,which was named as Cbuq PBP1. Bioinformation method was used to analyze its sequence signature and real-time PCR technology was used to measure the expression of Cbuq PBP1 in C. buqueti in different instars and tissues.【Results】The pheromone binding protein of C. buqueti was cloned and named as Cbuq PBP1(Gen Bank accession No:KU845733.1),which had open reading frame as 432 bp,143 encoded amino acid residues,molecular weight as 15.84 k D,isoelectric point as 4.60 and 6 conervative cysteine sites. According to the system evolutionary tree,Cbuq PBP1 was quite similar to PBP of other insects. According to the result of real-time PCR,Cbuq PBP1 was expressed in male,female and larval individuals. The expression in the males was the highest(P<0.05).Meanwhile,the expression in antennas of the male was significantly higher than that in other tissues(P<0.05).【Conclusion】It has been shown that the expression of Cbuq PBP1 in C. buqueti was different in different instars and tissues.

关 键 词：长足大竹象 CbuqPBP1 基因克隆 表达分析 

分 类 号：S763.38[农业科学—森林保护学]