幽门螺杆菌全菌蛋白电泳影响因素的研究

本文比较研究了用反复冻融法及用含十二烷基磺酸衲(SDS)和β-巯基乙醇的样品缓冲液直接破碎法处理幽门螺旋菌对全菌蛋白电泳结果的影响,并观察了两种方法处理后的菌体裂解情况。实验表明,反复冻融法破碎细菌不够彻底,经离心后有较多的膜性结构被清除,引起膜性结构与胞浆内成份不均匀损失,对全菌蛋白电泳条带(特别是某些条带的含量)有明显的影响。提示在作细菌全菌蛋白电泳,特别是用于进行定量分析时,不宜采用冻融法处理细菌,可考虑用样品缓冲液直接破碎细菌。超声波破碎及压力破碎法是否存在对电泳结果的类似影响有待进一步研究。

The Comparative Study of the Two Cells Lysing Methods on SDS-PAGE of Helicobacter Pylori

The comparative study was developed on the two methods of lysis on the SDS-PAGE,by repeated thawing and freezing and by the sam- ple buffer that contained SDS and 2-mercaptoethanol.Experiments showed when the HP cells were lysed by sample buffer,the cells were broken thoroughly,no pieces of HP cells were left,and about 40 protein bands were observed clearly.But when the cells were lysed by repeated thawing and freezing,pieces of HP cells were seen under the microscope.Some bands of protein became weak,only about 20 bands can be observed clearly.It is suggested that the repeated thawing and freezing method of lysis can affect the results of the SDS-PAGE, especially when the SDS-PAGE was used to analysing the protein amount and making the hierarchical clustering analysis.Whether the methods of press pouch and ultrasonification have similar effects on SDS-PAGE of HP whole cell proteins needs further investigation.