Objective To investigate the optimal standing time during Streptococcus mutans (one of caries-related bacterium) proteins extraction after the cells broken by ultrasonic. Methods S. mutans splitted under ultrasonic after suspending in cell lysis buffer. Contrast the forms of S. mutans before and after ultrasonic splitting. And the proteins were collected after bacterio-suspen-sion were standing for 0,10,30,50,70 min. Then the influence of standing time on extraction efficiency was studied by using SDS-PAGE and measuring the proteins concentration. Results Few cells broke incompletely after splitting. The protein concentration after standing for 30 min was higher than that of the others(P<0. 05). While the result of SDS-PAGE showed the abundance and distribution of protein bands were basically the same. It s supposed that the release and solubility of protein need necessary time to complete. Conclusion Standing for 30 min after S. mutans cells splitting could raise the proteins extraction efficiency.%目的 寻找超声破碎后高效率提取致龋菌变异链球菌蛋白质的最佳放置时间.方法 以裂解液为提取介质,采用超声法破碎细菌.对比破碎前、后菌形态并将菌提取液分别放置0、10、30、50、70 min收集蛋白上清液,通过RC DC法测定蛋白质浓度及蛋白质SDS-PAGE电泳,比较放置不同时间后的蛋白质提取效果.结果 超声破碎后,少部分细菌破碎不完全.RC DC法测定结果显示,超声破碎后放置30 min时提取的蛋白质浓度高于其他各组(P<0.05),但SDS-PAGE电泳显示蛋白质的条带分布和丰度无明显差别,推测蛋白质的释放和溶解需要一定时间完成.结论 超声法提取变异链球菌蛋白质后放置30 min可显著提高蛋白质提取效率.
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