葱蝇非滞育与夏滞育蛹蛋白的双向凝胶电泳比较分析

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

【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。

[ 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-methenyhetrahydrofolate 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.