›› 2008, Vol. 51 ›› Issue (7): 689-693.

• 研究论文 • 上一篇    下一篇

中华蜜蜂信息素结合蛋白ASP1 cDNA的克隆及时空表达


  • 出版日期:2008-07-20 发布日期:2008-07-20
  • 通讯作者: 高其康

Cloning and spatio temporal expression of cDNA encoding pheromone binding protein ASP1 in Apis cerana cerana (Hymenoptera: Apidae)

LI Hong-Liang   

  • Online:2008-07-20 Published:2008-07-20

摘要: 信息素结合蛋白(pheromone binding proteins, PBPs)在昆虫信息素的识别、传递和处理过程中具有重要作用。本研究首次克隆了中华蜜蜂Apis cerana cerana的一个PBP基因Ac-ASP1(GenBank序列号为DQ449670),其预测蛋白具有典型的气味结合蛋白(OBPs)标志(即成熟肽含有6个保守的半胱氨酸)。利用real-time PCR技术对Ac-ASP1在中蜂不同组织和发育历期的时空表达谱进行了鉴定。绝对定量结果显示Ac-ASP1高丰度地表达于工蜂触角(2.07×106 拷贝数/μg),而在其他组织(如头、胸、腹、翅及足)中呈低丰度表达(102拷贝数/μg); 相对定量结果显示Ac-ASP1在各发育历期如幼虫、蛹以及成虫发育早期(1~6日龄)均有大量表达,而在21日龄前后具有另外一个高丰度表达时期。这些结果可为明确Ac-ASP1在中蜂蜂王信息素信号识别传递过程中的作用提供参考。

关键词: 中华蜜蜂, 工蜂, 信息素结合蛋白, 克隆, 时空表达, 实时定量PCR

Abstract: Pheromone binding proteins (PBPs) play an important role in recognition, transmission and processing of insect pheromones. In this study, a cDNA named Ac-ASP1 encoding a PBP was cloned (from the Chinese native honeybee, Apis cerana cerana. The predicted protein has six conservative cysteines in mature peptide, a typical symbol of odorant binding proteins (OBPs). The sequence had been submitted into GenBank and registered is the accession no. DQ449670. The profile of spatio-temporal expression of Ac-ASP1 was identified by real-time PCR. The results of absolute quantitation showed that Ac-ASP1 was expressed at a higher level in antenna (2.07×106 copy number/μg), but at a less level in other tissues including head, thorax, abdomen, wings and legs (10.2 copy number/μg); while the results of relative quantitation revealed that Ac-ASP1 was expressed in two high abundance periods: from larva to the 6th day of adult and around the 21st day of adult. These results may provide the reference to ascertain the function of Ac-ASP1 involved in signal recognition and transmission of queen pheromone.

Key words: Apis cerana cerana, worker bee, pheromone binding proteins (PBPs), cloning, spatio-temporal expression, real-time PCR