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脆弱拟杆菌脂多糖的化学与免疫化学分析

Chemical and immunochemical analyses of Bacteroides fragilis lipopolysaccharides.

作者信息

Weintraub A, Larsson B E, Lindberg A A

出版信息

Infect Immun. 1985 Jul;49(1):197-201. doi: 10.1128/iai.49.1.197-201.1985.

Abstract

Lipopolysaccharides (LPSs) from 17 different Bacteroides fragilis strains were extracted in a two-step procedure. The first step was a hot phenol-water extraction of whole bacteria, resulting in a crude aqueous phase, which after lyophilization in a second step was extracted with a phenol-chloroform-light petroleum mixture. The resulting LPSs, which were essentially free from contaminating nucleic acid, proteins, and capsular polysaccharide, were investigated for their qualitative and quantitative sugar and fatty acid composition, immunochemical specificity by enzyme-linked immunosorbent inhibition assays, and particle weight by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Of the 17 strains, 13 had LPSs which all contained L-rhamnose, D-glucose, D-galactose, and D-glucosamine in approximately the same ratio. Three of these LPSs also contained D-galactosamine. The fatty acid composition was also similar in that the same fatty acids, although in slightly varying proportions, were found in all LPSs. The 13 strains also showed the same specificity in inhibition studies by enzyme immunoassay with rabbit anti-LPS antisera and LPS antigen. The LPS particle weights were also very similar, in the range of what is found for LPSs from rough mutant strains of enterobacteria. Our results suggest that most strains of B. fragilis have similar, if not identical, LPSs with relatively short polysaccharide chains.

摘要

采用两步法提取了17种不同脆弱拟杆菌菌株的脂多糖(LPS)。第一步是对全菌进行热酚 - 水提取,得到粗水相,第二步冻干后,用酚 - 氯仿 - 轻质石油混合物进行提取。对所得基本不含污染核酸、蛋白质和荚膜多糖的LPS,研究了其糖和脂肪酸的定性及定量组成、通过酶联免疫吸附抑制试验测定的免疫化学特异性以及通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳测定的颗粒重量。在这17个菌株中,有13个菌株的LPS均含有大致相同比例的L - 鼠李糖、D - 葡萄糖、D - 半乳糖和D - 葡糖胺。其中3种LPS还含有D - 半乳糖胺。脂肪酸组成也相似,即所有LPS中都含有相同的脂肪酸,尽管比例略有不同。在使用兔抗LPS抗血清和LPS抗原进行的酶免疫测定抑制研究中,这13个菌株也表现出相同的特异性。LPS颗粒重量也非常相似,处于肠道杆菌粗糙突变菌株LPS的重量范围内。我们的结果表明,大多数脆弱拟杆菌菌株具有相似(如果不是完全相同)的LPS,其多糖链相对较短。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76e3/262078/23f0377a21d9/iai00112-0207-a.jpg

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