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棉铃虫围食膜蛋白的电泳分离技术
Electrophoretic techniques for the separation of peritrophic membrane proteins from Helicoverpa armigera
李怡萍1**,仵均祥1,梁革梅2,郭予元2,袁向群1
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DOI:
作者单位:1西北农林科技大学植物保护学院 应用昆虫学重点实验室杨凌712100; 2中国农业科学院植物保护研究所 植物病虫害生物学国家重点实验室北京100193
中文关键词:棉铃虫, 围食膜蛋白, 分离, 电泳技术
英文关键词:Helicoverpa armigera, peritrophic membrane protein, separation, electrophoretic techniques
中文摘要:比较了棉铃虫Helicoverpa armigera (Hübner)围食膜不同处理方法,不同围食膜数量,在不同电泳下的分离效果,筛选最佳的围食膜蛋白分离技术。结果表明:冷冻干燥与非冷冻干燥处理围食膜后进行SDSPAGE电泳分离效果基本相同,建议使用冷冻干燥后处理较好。取1、3、5条围食膜进行SDSPAGE电泳,都能将围食膜蛋白分离,且分离效果基本一致,建议5条围食膜最合适,具有可靠性和代表性。浓缩胶为5%、分离胶分别为8%、10%和12%时棉铃虫围食膜的SDSPAGE电泳分离效果差异不大,而分离胶为12%时效果较好。无水三氟利克酸处理围食膜后进行NuPAGE电泳,分离出的围食膜蛋白大约30多种,远远超过上述方法分离的16~20种,且需要的围食膜材料少,分离效果最佳,但费用高。
英文摘要: The effects of the separation of peritrophic membrane (PM) proteins from Helicoverpa armigera (Hübner)were compared using different treatment methods, different numbers of PM proteins and different electrophoretic conditions, in order to select the optimal technique. The results of SDSPAGE show that, although freezedrying is recommended for the separation of PM proteins, nearly equivalent results were achieved with and without freeze drying. Although one, three and five strips of PM protein could be completely separated by SDSPAGE, five strips provide a more reliable representation of the population. There was no significant difference in the results obtained by SDSPAGE using separation gel concentrations of 8%、10% and 12% and a stacking gel concentration of 5%. The optimum separation gel concentration was 12%. Treating PM with anhydrous trifluoromethanesulfonic acid before separation with NuPAGE resulted in approximately 30 types of PM protein, far exceeding the 16-20 types of proteins previously obtained from H. armigera. Furthermore, only a small amount of PM material was needed using this method. However, although this method was the most effective it is considerably more expensive than the alternatives.
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