›› 2014, Vol. 57 ›› Issue (2): 161-167.

• 研究论文 • 上一篇    下一篇

葱蝇非滞育与夏滞育蛹蛋白的双向凝胶电泳比较分析

郝友进, 胡文霞, 陈斌*   

  1. (重庆师范大学生命科学学院, 昆虫与分子生物学研究所, 重庆 401331)
  • 出版日期:2014-02-20 发布日期:2014-02-20

Comparative analysis of proteins in non-and summer-diapausing pupae of the onion fly, Delia antiqua (Diptera: Anthomyiidae), using two-dimensional gel eletrophoresis

HAO You-Jin, HU Wen-Xia, CHEN Bin*   

  1. (Institute of Entomology and Molecular Biology, College of Life Sciences, Chongqing Normal University, Chongqing 401331, China)
  • Online:2014-02-20 Published:2014-02-20

摘要: 【目的】比较分析葱蝇Delia antiqua非滞育与夏滞育蛹的蛋白表达差异, 为进一步揭示昆虫滞育的分子调控机理和昆虫防治提供理论基础。【方法】以非滞育和夏滞育的蛹为材料, 提取总蛋白; 进行双向凝胶电泳和凝胶图像分析, 对并差异蛋白质进行MALDI-TOF MS质谱鉴定, 获得该点的质量指纹图谱; 利用MASCOT软件在NCBI和SWISS PORT蛋白质数据库中进行搜索鉴定。【结果】非滞育和夏滞育蛹的蛋白表达存在显著差异。通过质谱鉴定和生物信息学分析, 13个差异表达的蛋白质分别为胶原蛋白、 纺锤丝组装非正常蛋白6(SAS6)、 5,10-亚甲基四氢叶酸合成酶(MTHFS)、  Bnb蛋白(bangles and beads)及其他功能未知蛋白。【结论】葱蝇在夏滞育时期, 蛹体内的某些蛋白被上调或下调表达。本研究所鉴定的蛋白中, 部分可能是参与滞育相关的蛋白质网络中的成员, 它们可能在抗高温、 染色体分离、 叶酸代谢等生理过程中发挥重要作用。

关键词: 葱蝇, 滞育, 总蛋白, 双向凝胶电泳, 质谱分析, 生物信息学分析

Abstract: 【Aim】 Comparative analysis was performed to investigate differentially expressed proteins between non- and summer-diapausing pupae of the onion fly, Delia antiqua, and to provide a theoretical basis for further exploring the molecular regulation mechanisms of insect diapause and pest control management. 【Methods】 Total proteins were extracted from non-and summer-diapausing pupae, separated by two-dimensional gel electrophoresis, and then analyzed by image analysis. Peptide mass fingerprinting of differential proteins were analyzed by MALDI-TOF MS and identified through database searching in NCBI and SWISS PORT using MASCOT. 【Results】 Proteins were differentially expressed in non- and summer-diapausing pupae. Thirteen differentially expressed proteins were identified by mass spectrometry and bioinformatic analysis, including collagen, SAS6 (spindle assembling abnormal protein 6), MTHFS (5, 10-methenyltetrahydrofolate synthetase), Bnb (bangles and beads) and proteins with unknown functions. 【Conclusion】 The expressions of some proteins are up- or down-regulated in summer-diapausing pupae. Proteins identified in this study may be members of diapause-related protein regulatory networks, and they potentially play important roles in high temperature resistance, chromosome separation, folate metabolism and other physiological processes. 

Key words: Delia antiqua, diapause, total protein, two-dimentional gel electrophoresis, mass spectrographic analysis, bioinformatic analysis