伤寒沙门菌野生型与粗糙型菌体蛋白质的双向电泳分析与研究

Analysis and research of the proteins for the wild type and the rough type of Salmonella typhi by 2D-PAGE

了解伤寒沙门菌野生型与粗糙型菌株的菌体蛋白质组成特点，探讨伤寒沙门菌粗糙型变异的遗传学基础。分离提取伤寒沙门菌野生型与粗糙型菌株的菌体蛋白质，聚丙烯酰胺凝胶双向电泳（2D-PAGE）和考马斯亮蓝染色，计算机分析与比较两菌株的蛋白质组成特点及其相关性。伤寒沙门菌野生型与粗糙型具有相似的蛋白质电泳图谱，相似系数为78％。多数蛋白质斑点分布于pH3．0～6．4之间，并且分子量小于30kDa。两菌株的蛋白质电泳图谱之间的主要差异共计36处，多数差异蛋白质的分子量小于20kDa。伤寒沙门菌粗糙型与野生型菌体蛋白质组成的差异显示，粗糙型变异绝不仅仅是O抗原多糖的缺失，也可发生菌体蛋白质组成的改变与缺失。伤寒沙门菌粗糙型保留了同其亲代野生型菌株一致的绝大多数菌体蛋白质组成，在2D-PAGE中形成伤寒沙门菌特征性的基本蛋白质图谱，有助于对粗糙型菌株进行蛋白质分子同源性与变异性的分析与鉴别。

To find out the protein properties of the wild type and the rough type of Salmonella typhi （ S. typhi） ; to probe the genetic mechanism of the rough variation of S, typhi. The proteins were separated from the wild type and the rough type of S. typhi, and then the samples were processed by 2D-PAGE and stained by Coomassie Brilliant R-250. The properties and relationship of the protein patterns were analyzed by the software hnagemaster 6.0. By the 2D-PAGE, the sinfilar protein patterns with 78% Sd （similar data） would be found between the wild type and the rough type of S. typhi. The distribution of most protein spots was in the range of pH 3.0 - 6.4 where the molecular weight of protein was smaller than 30kDa. According to the characters of differential proteins, there is not only the polysaccharide side chain of the O antigen but also some proteins that could be lost or changed in the rough strain of S. typhi. Most proteins in it would be the same as those in the parental bacteria so that the characteristic protein pattern of Salmonella can be found by 2D-PAGE. This property is useful for analysis and identification of the homologue between variations and their parental bacteria.