PROCEDURE FOR ISOLATION OF BACTERIAL LIPOPOLYSACCHARIDES FROM BOTH SMOOTH AND ROUGH PSEUDOMONAS-AERUGINOSA AND SALMONELLA-TYPHIMURIUM STRAINS
PROCEDURE FOR ISOLATION OF BACTERIAL LIPOPOLYSACCHARIDES FROM BOTH SMOOTH AND ROUGH PSEUDOMONAS-AERUGINOSA AND SALMONELLA-TYPHIMURIUM STRAINS
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DOI：
10.1128/jb.155.2.831-838.1983
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发表时间：
1983-01-01
影响因子：
3.2
通讯作者：
HANCOCK, REW
中科院分区：
文献类型：
--
作者：
DARVEAU, RP;HANCOCK, REW
脂多糖（LPS）是革兰氏阴性菌外膜的主要成分。在单个生物体内，这种分子的大小异质性可能存在。开发了一种LPS分离方法，该方法可有效地以高产率（通过使用羟基脂肪酸、庚糖和2-酮-3-脱氧辛酸酯产率定量，存在于全细胞中的LPS的51-81%）和高纯度提取光滑和粗糙LPS。蛋白质（0.1%重量的LPS）、核酸（1%）、脂质（2-5%）和其它细菌产物的污染较低。LPS的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳证明了在分离的光滑LPS中存在高度的大小异质性，并且存在大量的粗糙型LPS。在多种免疫化学分析中，铜绿假单胞菌LPS与单克隆抗体良好地相互作用。通过比较从铜绿假单胞菌和S.鼠伤寒。铜绿假单胞菌的抗生素超敏感突变体Z61的LPS实际上仅含有小比例的O-抗原性侧链，该突变体先前在通过苯酚-水程序分离的光滑LPS分子与粗糙LPS分子的比率方面被表征为与野生型相同。
Lipopolysaccharide (LPS) is a major component of the outer membrane of gram-negative bacteria. Within a single organism, size heterogeneity of this molecule can exist. A LPS isolation procedure which is effective in extracting smooth and rough LPS in high yields (51-81% of the LPS present in whole cells as quantitated by using hydroxy fatty acid, heptose and 2-keto-3-deoxyoctonate yields) and with a high degree of purity was developed. The contamination by protein (0.1% by weight of LPS), nucleic acids (1%), lipids (2-5%) and other bacterial products was low. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the LPS demonstrated the presence of a high degree of size heterogeneity in the isolated smooth LPS and the presence of significant amounts of rough-type LPS. The P. aeruginosa LPS interacted well with a monoclonal antibody in a variety of immunochemical analyses. The usefulness of the procedure was demonstrated by comparing LPS preparations obtained from wild-type and mutant strains of P. aeruginosa and S. typhimurium. The LPS of an antibiotic supersusceptible mutant Z61 of P. aeruginosa, which was previously characterized as identical to wild type with respect to the ratio of smooth to rough LPS molecules isolated by the phenol-water procedure, actually contained only a small proportion of O-antigenic side chains.