›› 2018, Vol. 36 ›› Issue (01): 78-85.

• • 上一篇    下一篇

长足大竹象信息素结合蛋白CbuqPBP1克隆和表达分析

杨桦   

  1. 贵州大学昆虫研究所;四川农业大学森林保护重点实验室
  • 收稿日期:2017-05-18 修回日期:2018-01-23 出版日期:2018-02-28 发布日期:2018-11-29
  • 通讯作者: 杨桦 E-mail:yanghua151017@163.com
  • 基金资助:
    中国博士后科学基金面上资助项目;国家自然科学基金

Molecular cloning and expression analysis of CbuqPBP1 gene in the bamboo snout beetle, Cyrtotrachelus buqueti

YANG /HUA   

  • Received:2017-05-18 Revised:2018-01-23 Online:2018-02-28 Published:2018-11-29
  • Contact: YANG /HUA E-mail:yanghua151017@163.com
  • Supported by:
    ;National Natural Science Foundation of China

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,命名为CbuqPBP1。采用生物信息方法分析了其序列特征,利用实时荧光定量PCR技术研究了CbuqPBP1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CbuqPBP1(GenBank登录号:KU845733.1),开放阅读框为432 bp,编码143个氨基酸残基,分子量为15.84kD,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,CbuqPBP1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,CbuqPBP1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CbuqPBP1在长足大竹象不同虫态和不同组织中差异表达。

关键词: 长足大竹象, CbuqPBP1, 基因克隆, 表达分析

Abstract: 【Objective】This study aims to clone the pheromone binding proteins (PBPs) of the bamboo snout beetle Cyrtotrachelus buqueti Guerin-Meneville, and to analyze its gene sequence and expression profiles, so as to further research the physiological function of this gene. 【Methods】The reverse transcription PCR (RT-PCR) method was used to amplify PBP (designate as CbuqPBP1). Bioinformatic methods and real-time fluorescence quantitative PCR were used to analyze the characteristics of CbuqPBP1 and relative expression level in different stages and different tissues. 【Results】The results showed that a PBP cloned from C. buqueti, which was named CbuqPBP1 (GenBank accession No:KU845733.1), the sequence of CbuqPBP1 contains a 432 bp open reading frame that encodes 143 amino acids, with a molecular mass of 15.84KD, and an isoelectric point of 4.60. The phylogenetic tree showed that CbuqPBP1 has high comparability with the PBP of other insects. The results of real-time fluorescence quantitative PCR showed that CbuqPBP1 was expressed in the body of male, female and larval, the highest expression level was in the body of male (P<0.05). Meanwhile, the expression level in male antennae was significantly higher than other tissues (P<0.05). 【Conclusion】Manifest that CbuqPBP1 has different expression in different insect states and different tissues of C. buqueti.

Key words: Cyrtotrachelus buqueti, CbuqPBP1, gene cloning, expression analysis

中图分类号: 

  • Q968