›› 2009, Vol. 52 ›› Issue (2): 140-146.

• 研究论文 • 上一篇    下一篇



  • 出版日期:2009-03-18 发布日期:2009-02-20
  • 通讯作者: 王满囷

Molecular cloning and expression characterization of a chemosensory protein gene PxylCSP1 from the diamondback moth, Plutella xylostella (Lepidoptera: Plutelidae)

  • Online:2009-03-18 Published:2009-02-20

摘要: 利用RT-PCR和RACE技术克隆到小菜蛾Plutella xylostella 化学感受蛋白(CSP)基因PxylCSP1(GenBank登录号: FJ361903),其核苷酸序列全长405 bp,编码134个氨基酸残基,预测N-末端包含19个氨基酸组成的信号肽序列,估测其成熟蛋白分子量为13.56 kD,等电点为6.12。该基因编码氨基酸序列和其他鳞翅目昆虫CSP的氨基酸序列比对同源性较高(70%~80%)。RT-PCR结果表明PxylCSP1不仅存在于小菜蛾的触角中,还存在于头、足、腹和翅中。Real-time PCR结果表明PxylCSP1的表达水平因被测小菜蛾的性别、日龄、组织不同和交配与否而异。

关键词: 小菜蛾, 化学感受蛋白, 分子克隆, 反转录聚合酶链式反应, 实时定量聚合酶链式反应

Abstract: We described the identification and characterization of a new cDNA encoding chemosensory protein1 (CSP1) from the antenna of diamondback moth, Plutella xylostella, a species whose CSPs have not been identified to date. We focused our investigation on this olfactory protein family using reverse transcription-polymerase chain reaction strategies. The results showed that the CSP1 gene in diamondback moth, named PxylCSP1 (GenBank accession no. FJ361903), is 405 bp in length and encodes 134 amino acid residues, with six cysteine residues in conserved positions relative to other known CSPs. The predicted N-terminus hydrophobic region contains 19 amino acid residues within the Has-CSP, displaying the characteristic features of a signal peptide. Thus the predicted molecular weight (MW) of the mature protein is 13.56 kD and isoelectric point (pI) is 6.12. The alignment of PxylCSP1 showed high sequence identity (70%-80%) with other full-length sequences of other lepidoteran CSPs from GenBank. RT-PCR analysis revealed that PxylCSP1 was not only expressed in antennae, but also in head, abdomen, wings and legs. The reasult of real-time PCR further indicated that the transcription level of PxylCSP1 depended on the gender, age, mating status and tissues of the tested moths.

Key words: Plutella xylostella, chemsensory proteins, molecular cloning, RT-PCR, real-time PCR