水貂绿脓杆菌（PASD03株）鞭毛蛋白的提取与分析

本试验以水貂绿脓杆菌PASD03株为研究对象,对其鞭毛蛋白的提取方法进行研究,分别通过酸裂解、差速离心以及酸裂解结合机械振荡三种方法,用（NH4）2SO4盐析来获得鞭毛蛋白,采用垂直板聚丙烯酰胺凝胶电泳（SDS-PAGE）对提取的绿脓杆菌鞭毛蛋白进行分离纯化,并进行电镜鉴定。结果显示：成功分离出水貂绿脓杆菌的鞭毛蛋白,获得了绿脓杆菌鞭毛蛋白的初提液.绿脓杆菌鞭毛蛋白经SDS-PAGE提示纯化的鞭毛蛋白为一条相对分子质量（Mr）为53×103蛋白带,透射电镜（SEM）观察发现该鞭毛蛋白呈丝状。结论：经酸裂解法并结合机械振荡和硫酸铵沉淀法简便、耗时短、纯度高、产量高,适用于绿脓杆菌鞭毛蛋白的提取。

Extraction and Analysis of FlageUin from Pseudomonas Aeruginosa

In the study, the extraction of flagellin from Pseudomonas aeruginosa（Strain PASD03 ）was studied, Flagellin was prepared through acid hydrolysis, differential centrifugation and acid hydrolysis in combination with mechanical oscillation.The flagellar protein structure from Pseudomonas aeruginosa was separated and purified by means of SDS-PAGE, and identified by electronic microscopy. The results showed that the experiment had successfully separated the flagellin of Pseudomonas aeruginosa and the original flagellin extract was obtained with a 53kD band of flagellin in SDS-PAGE. The purified flagellin showed the typical filamentous under electronic microscopy.Conclusion： The acid hydrolysis combined with mechanical oscillation and ammonia sulfate precipitation method was suitable for flagellin extraction because the simple and time-saving method could provide flagellin with high purity and high yield.