›› 2008, Vol. 51 ›› Issue (12): 1304-1308.

• 研究论文 • 上一篇    下一篇

烟夜蛾G蛋白αq亚基(HassGαq)的抗血清制备及在触角中的免疫定位

乔奇,李海超,原国辉,郭线茹,罗梅浩   

  • 出版日期:2010-07-29 发布日期:2008-12-20
  • 通讯作者: 原国辉

Preparation of the antiserum to HassGαq of Helicoverpa assulta(Lepidoptera: Noctuidae) and its immunolocalization in the antenna

QIAO Qi   

  • Online:2010-07-29 Published:2008-12-20

摘要: 为了明确烟夜蛾Helicoverpa assulta G蛋白αq亚基(HassGαq)在雄性触角中的定位,进而探索该亚基在烟夜蛾嗅觉信号传导过程中的作用,本实验首先以原核表达的HassGαq蛋白作为抗原免疫Balb/c小鼠,制备了抗血清。SDS-PAGE分离雄性烟夜蛾触角匀浆上清液后,Western blot检测结果表明,所制备的抗血清可识别HassGαq蛋白,同时也与非目的蛋白发生反应,说明在雄性烟夜蛾触角中HassGαq基因可能有多个转录本,或者可能有多种G蛋白α亚基表达。此外,商品化的anti-Gq/11α抗血清可特异地识别HassGαq蛋白。因此,利用anti-Gq/11α抗血清和免疫组化方法对HassGαq蛋白在雄性烟夜蛾触角中进行定位,结果显示HassGαq在雄性烟夜蛾触角毛形感器和锥形感器的基部以及感器腔中皆有表达。据此推测HassGαq可能参与烟夜蛾的嗅觉信号传导。

关键词: 烟夜蛾, G蛋白αq亚基, 抗血清, 免疫定位 嗅觉信号传导

Abstract: The objective of this study is to localize G protein αq subunit of Helicoverpa assulta (HassGαq) in the male antennae for the further research about the role of HassGαq in olfactory signal transduction in H. assulta. Balb/c mice were immunized with HassGαq protein expressed in prokaryotic cells to produce antiserum. The prepared antiserum and the purchased anti-Gq/11α antiserum were used in Western blot after male antennal homogenate of Helicoverpa assulta was separated by SDS-PAGE. The results indicated that the prepared antiserum could react with HassGαq, and also the non-target proteins, suggesting that several transcripts of HassGαq or other G protein α subunits may be expressed in male antennae. In addition, anti-Gq/11α antiserum could detect HassGαq with high specificity. Accordingly, the expression sites of HassGαq in adult male antennae were examined by immunocytochemical method using anti-Gq/11α antiserum. The results revealed that HassGαq was associated with trichodic sensilla and basiconic sensilla as immunolabelling were typically observed inside and at the base of the two kinds of sensilla. It is so inferred that HassGαq may be involved in olfactory signal transduction in H. assulta.

Key words: Helicoverpa assulta, G protein αq subunit, antiserum, immunolocalization, olfactory signal transduction