Emel'ianov V V, Balaeva N M
Mol Gen Mikrobiol Virusol. 1990 Apr(4):28-32.
The methods of cell lysis by lysozyme in tris-EDTA-sucrose with the consequent disruption of spheroplasts by the osmotic shock were used to obtain the total membranes from the intact or temperature-inactivated Rickettsia prowazekii. Detergents solubilization methods were used for analysis of outer membrane proteins. Sarcosyl insoluble material is shown to contain the main 134, 31, 29.5 and 25 Kd proteins, the minor 78, 60, 42, 17 Kd proteins, while the mixture of both membranes possess a more complex composition. Treatment of total membranes by the 2% octylglycoside results in elimination of the 31 Kd polypeptide. Inactivated Rickettsia can be used for isolation of the outer layer proteins diminishing the risk of working with this pathogenic microorganism.
采用溶菌酶在三羟甲基氨基甲烷-乙二胺四乙酸-蔗糖溶液中进行细胞裂解,随后通过渗透休克破坏原生质球的方法,从完整的或经温度灭活的普氏立克次氏体中获取总膜。使用去污剂溶解方法分析外膜蛋白。显示十二烷基肌氨酸钠不溶性物质含有主要的134、31、29.5和25千道尔顿蛋白、次要的78、60、42、17千道尔顿蛋白,而两种膜的混合物具有更复杂的组成。用2%辛基糖苷处理总膜会导致31千道尔顿多肽的消除。灭活的立克次氏体可用于分离外层蛋白,从而降低使用这种致病微生物的风险。