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目的探索制备灭蚊丝状真菌Pythiumsp.GY1938菌株菌丝体可溶性蛋白图谱的最佳染色方案。方法采用不同的破壁方法制备Pythiumsp.GY1938菌丝体可溶性蛋白,并通过SDS-PAGE进行蛋白质电泳分离,进一步采用考马斯亮蓝R-250染色法、Vorum银染法、EMBL银染法、Blum银染法4种染色方案分别进行凝胶染色,对不同染色方案的染色效果进行分析和比较。结果对于Pythiumsp.GY1938菌丝体可溶性蛋白样品中的中、高分子质量蛋白(≥44ku),考马斯亮蓝R-250的染色效果较好,且简单、快捷;对于低分子质量(<44ku)的可溶性蛋白,Vorum银染法染色效果较好;Blum银染法和EMBL银染法的染色效果均不理想。结论考马斯亮蓝R-250染色法和Vorum银染法均可应用于Pythiumsp.GY1938菌丝体可溶性蛋白图谱的制备,前者适合对中、高分子质量蛋白染色,后者适合对低分子质量蛋白染色。
Objective To explore the best staining scheme for preparing the mycelium soluble protein map of Pythium sp.GY1938, an antifungal filamentous fungus. Methods The mycelium soluble protein of Pythium sp. GY1938 was prepared by different methods of breakage. The protein was separated by SDS-PAGE and further purified by Coomassie brilliant blue R-250, Vorum silver staining, EMBL silver staining, Blum silver Dyeing four kinds of staining programs were gel staining, staining of different staining programs were analyzed and compared. Results The staining results of Coomassie Brilliant Blue R-250 were good and simple and fast for medium and high molecular weight proteins (≥44ku) in Pythium sp. GY1938 mycelium soluble protein samples. For low molecular mass (<44ku) Soluble protein, Vorum silver staining staining better; Blum silver staining and EMBL silver staining staining results are not satisfactory. Conclusion Coomassie brilliant blue R-250 and Vorum silver staining can be applied to the preparation of Pythium sp. GY1938 mycelium soluble protein profiles, the former suitable for medium and high molecular weight protein staining, the latter suitable for low molecular weight protein staining .