SUPRAMOLECULAR STRUCTURE OF RIGID LAYER OF CELL WALL OF SALMONELLA, SERRATIA, PROTEUS, AND PSEUDOMONAS-FLUORESCENS - NUMBER OF LIPOPROTEIN MOLECULES IN A MEMBRANE LAYER
SUPRAMOLECULAR STRUCTURE OF RIGID LAYER OF CELL WALL OF SALMONELLA, SERRATIA, PROTEUS, AND PSEUDOMONAS-FLUORESCENS - NUMBER OF LIPOPROTEIN MOLECULES IN A MEMBRANE LAYER
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DOI：
10.1021/bi00828a001
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发表时间：
1970-01-01
期刊：
影响因子：
2.9
通讯作者：
WOLFF, H
中科院分区：
文献类型：
--
作者：
BRAUN, V;REHN, K;WOLFF, H
材料与方法菌株与培养条件。使用以下菌株：鼠伤寒沙门氏菌LT 2、沙门氏菌mR 608和沙门氏菌usumbura（均获自O. Liideritz，弗赖堡）、粘质沙雷氏菌、奇异变形杆菌ATCC 2505，其需要胸腺嘧啶和烟酸并且是链霉素抗性的（D. Helinsky，拉霍亚）、奇异变形杆菌VI和1 Ca/5（H. Martin，达姆施塔特）、普通变形杆菌ATCC 13315和荧光假单胞菌。除非另有说明（参见表IV的变形杆菌属），细胞在Difco培养基3中生长，该培养基根据菌株的要求补充有另外的生长因子; 2.5-1.用10-ml过夜培养物接种瓶，并在37 ℃下剧烈通气，细胞生长至约5 × 105个细胞/ml的密度。将细胞用冰冷却，离心，并用水洗涤一次;获得3- 5g细胞。在16-1中增长了更大的数量。瓶子。刚性层的隔离。用玻璃珠振摇细胞，然后如所述进行差速离心（Braun和Sieglin，1970）。用0.01mEDTA，pH 8洗涤由细胞膜组成的沉淀一次，然后悬浮在水中，滴加到沸腾的4%十二烷基硫酸钠（W. Leutgeb，personal communication）.如所述（Braun和Rehn，1969），将不溶性刚性层离心并用水洗涤。用S的刚性层重复十二烷基硫酸钠处理。miniatum，S. usumbura，以及变形杆菌属和荧光假单胞菌属。
Materials and MethodsStrains and Culture Conditions. The following strains were used: Salmonella typhimurium LT2, Salmonella minnesota mR 608 and Salmonella usumbura (both obtained from O. Liideritz, Freiburg), Serratia marcescens, Proteus mirabilis ATCC 2505 which required thymine and nicotinic acid and which was streptomycin resistent (D. Helinsky, La Jolla), Proteus mirabilis VI and 1 Ca/5 (H. Martin, Darmstadt), Proteus vulgaris ATCC 13315, and Pseudomonas fluorescens. Unless otherwise indicated (see Table IV for Proteus), the cells were grown in Difco antibioticmedium 3 supplemented with additional growth factors according tothe requirements of the strains; 2.5-1. bottles were inoculated with 10-ml overnight cultures and vigorously aeratedat 37, and the cells grown to a density of about 5 X 10s cells/ml. The cells were chilled with ice, spun down, and washed once with water; 3-5 g cells of was obtained. Larger quantities were grown in 16-1. bottles.Isolation of the Rigid Layer. Cells were shaken with glass beads followed by differential centrifugation as described (Braun and Sieglin, 1970). The pellet consisting of the cell en-velope was washed once with 0.01 m EDTA, pH 8, then suspended in water, and added dropwise into a boiling solution of 4% sodium dodecyl sulfate (W. Leutgeb, personal com-munication). The insoluble rigid layer was spun down and washed with water as described (Braun and Rehn, 1969). The sodium dodecyl sulfate treatment was repeated with the rigid layer of S. minnesota, S. usumbura, and Proteus and Pseudo-monas fluorescens.