论文部分内容阅读
目的探索建立有效的蜡样芽孢杆菌(Bacillus cereus)蛋白质组双向电泳体系,为进一步揭示其促进氧化葡萄糖酸杆菌(Gluconobacter oxydans)产酸的作用机制奠定基础。方法以培养的B.cereus为材料,比较蛋白质制备超声破壁时间、新型细胞裂解液、pH梯度和不同上样量对B.cereus蛋白双向电泳结果的影响。结果与结论采用15 min超声破壁提取B.cereus总蛋白,选用新型蛋白质裂解,用长24 cm、pH 4～7的IPG胶条,采用80μg上样量进行等电聚焦,于60 V 15 min、120 V 6 h条件下进行SDS-PAGE垂直电泳,可以获得背景清晰、重复性好的双向电泳图谱,建立一套用于B.cereus蛋白质组分析的双向电泳方法。
Objective To explore the establishment of an effective two-dimensional electrophoresis system of Bacillus cereus proteomics and lay a foundation for further revealing its mechanism of promoting acid production by Gluconobacter oxydans. Methods The cultured B.cereus was used as material to compare the effects of protein breaking time, new cell lysate, pH gradient and different loading on the two-dimensional electrophoresis results of B.cereus. RESULTS AND CONCLUSION: The total protein of B.cereus was extracted by sonication for 15 min. New protein was used for proteolysis. IPG strips of 24 cm in length and pH 4-7 were used for isoelectric focusing at 80 μg for 60 min at 15 min , 120 V 6 h under the conditions of SDS-PAGE vertical electrophoresis, you can get a clear background, good repeatability of the two-dimensional electrophoresis, set up a set for the B.cereus proteome analysis two-dimensional electrophoresis.