双向电泳分析链格孢霉过敏原提取液的致敏蛋白组分

Analysis of Alternaria aiternata allergen by 2-diamensional electrophoresis

目的利用双向电泳及其免疫印迹分析技术研究链格孢霉过敏原提取液的致敏组分。方法对经空气曝皿获得，经形态学和分子生物学鉴定为链格孢霉的真菌进行26％恒温下培养4周，提取新鲜菌苔总蛋白后，用双向电泳技术进行蛋白质分离，免疫印迹技术分析链格孢霉过敏原提取液致敏蛋白组分。结果链格孢霉过敏原提取液经双向电泳共发现1450±35个蛋白点，双向免疫印迹分析发现6种过敏原组分，它们的相对分子质量（M，）和等电点（PI）分别为：16．4×10^3，PI为7．0—8．0；26×10^3，PI为6．0～7．0；31×10^3，PI为7．5～8．5；34×10^3，PI为6．5～7．0；45×10^3，PI为7．0—8．0；57×10^3，PI为6．5～7．5。结论双向电泳可用于链格孢霉过敏原提取液致敏蛋白组分的分析。

Objective To explore the components of Ahernaria alternata allergen with two-dimen- sional electrophoresis. Methods The fungus was gotten from air, identified by morphology and molecular as Alternaria ahernata, cultured under 26℃ for 4 weeks, extracted total protein of fresh molds, isolated with two-dimensional electrophoresis. And the components of allergen were analyzed with Western blot. Results Two-dimensional electrophoresis showed 1450 ± 35 protein spots. Six allergen components were found by two- dimensional electrophoresis Western blot. Their molecular weight and PI were as follows： 16.4 ×10^3 , PI7.0- 8.0 ;26 ×10^3, PI6.0-7.0 ;31×10^3 , PI7.5-8.5 ; 34×10^3 , PI6.5-7.0 ;45×10^3 , PI7.0-8.0 ; 57 ×10^3 , PI6.5- 7.5. Conclusion Two-dimensional electrophoresis could be used in analyzing allergen of Alternaria ahernata.