The type and yield of lipopolysaccharide from symbiotically deficient rhizobium lipopolysaccharide mutants vary depending on the extraction method.
The type and yield of lipopolysaccharide from symbiotically deficient rhizobium lipopolysaccharide mutants vary depending on the extraction method.
复制标题
来自共生缺陷根瘤菌脂多糖突变体的脂多糖的类型和产量根据提取方法而变化。
DOI：
10.1093/glycob/10.10.1013
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发表时间：
2000
期刊：
影响因子：
4.3
通讯作者：
Carlson,RW
中科院分区：
文献类型：
--
作者：
Ridley,BL;Jeyaretnam,BS;Carlson,RW
至少有18种脂多糖（LPS）提取方法可用，没有一种方法是普遍适用的。本研究采用热酚/水（H2O/W）和酚/EDTA/三乙胺（H2O/EDTA/TEA）两种提取方法，从4株大豆根瘤菌、1株三叶根瘤菌24AR和大豆根瘤菌CE3中分离出了脂多糖。通过脱氧胆酸盐聚丙烯酰胺凝胶电泳（DOC-PAGE）和免疫印迹法对各种制剂中的LPS进行定量分析。这些根瘤菌通常具有两种突出的LPS形式：LPS I，其具有O-多糖，和LPS II，其没有。获得的LPS形式取决于提取方法，并根据提取的突变体而变化。两种方法都从CE3中提取LPS I和LPS II。EDTA/TEA法而不是EDTA/W法从突变体CE 358和CE 359中提取LPS I。相反地，ESTA/W而不是ESTA/EDTA/TEA方法提取CE359 LPS V，其为具有截短的0-多糖的LPS形式。突变体CE406的EDTA/TEA提取得到良好的LPS I和II产率，而EDTA/W提取得到非常少量的LPS I。所有的LPS产量 使用EDTA/EDTA/TEA提取的菌株的产率是相当一致的（3倍范围），而来自EDTA/W提取的产率是高度可变的（850倍范围）。EDTA/TEA方法从突变体24 AR中提取LPS I和LPS II，但EDTA/W方法将LPS II专门分配到苯酚相中，使其难以回收。总的来说， EDTA/TEA提取从突变体中产生更多形式的LPS，并提供了一种更简单、更快、危害更小的替代方法来代替EDTA/W提取。然而，可以得出结论，任何LPS突变体的仔细分析需要使用一种以上的提取方法。
At least 18 lipopolysaccharide (LPS) extraction methods are available, and no single method is universally applicable. Here, the LPSs from fourR.etli, oneR.leguminosarumbv.trifoliimutant, 24AR, and theR.etliparent strain, CE3, were isolated by hot phenol/water (ϕ/w), and phenol/EDTA/triethylamine (ϕ/EDTA/TEA) extraction. The LPS in various preparations was quantified, analyzed by deoxycholate polyacrylamide gel electrophoresis (DOC-PAGE), and by immunoblotting. These rhizobia normally have two prominent LPS forms: LPS I, which has O-polysaccharide, and LPS II, which has none. The LPS forms obtained depend on the method of extraction and vary depending on the mutant that is extracted. Both methods extract LPS I and LPS II from CE3. The ϕ/EDTA/TEA, but not the ϕ/W, method extracts LPS I from mutants CE358 and CE359. Conversely, the ϕ/W but not the ϕ/EDTA/TEA method extracts CE359 LPS V, an LPS form with a truncated O-polysaccharide. ϕ/EDTA/TEA extraction of mutant CE406 gives good yields of LPS I and II, while ϕ/W extraction gives very small amounts of LPS I. The LPS yield from all the strains using ϕ/EDTA/TEA extraction is fairly consistent (3-fold range), while the yields from ϕ/W extraction are highly variable (850-fold range). The ϕ/EDTA/TEA method extracts LPS I and LPS II from mutant 24AR, but the ϕ/W method partitions LPS II exclusively into the phenol phase, making its recovery difficult. Overall, ϕ/EDTA/TEA extraction yields more forms of LPS from the mutants and provides a simpler, faster, and less hazardous alternative to ϕ/W extraction. Nevertheless, it is concluded that careful analysis of any LPS mutant requires the use of more than one extraction method.