Izard Jacques, Limberger Ronald J
New York State Department of Health, Wadsworth Center, David Axelrod Institute for Public Health, P.O. Box 22002, Albany, NY 12201-2002, USA.
J Microbiol Methods. 2003 Nov;55(2):411-8. doi: 10.1016/s0167-7012(03)00193-3.
Although specific lipids in bacteria can be quantitated, there is still a need to quantitate the total lipid content of a bacterial sample. The sulfo-phospho-vanillin reaction for quantitation of bacterial lipids has significant advantages over traditional methods for screening of engineered mutant strains. In this report we show that this methodology can be used directly on whole cell or homogenized biological material, without any extraction step. The cell components, and most of the reagents used for cell extraction, that were tested did not interfere with the reaction. The screening is based on the observation of physiologic variations using ratios of relative amounts: lipid/DNA and lipid/protein. Our results show that significant differences in those ratios can be detected when there is a modification of the phospholipid content of the cell. The sample manipulation required is minimal and could be automated. Used as a primary screening and/or characterization of engineered mutant strain, the test may lead to further investigation of the nature and distribution of lipids in the cell.
尽管细菌中的特定脂质可以进行定量分析，但仍需要对细菌样本的总脂质含量进行定量。用于定量细菌脂质的磺基 - 磷 - 香草醛反应相对于传统的工程突变菌株筛选方法具有显著优势。在本报告中，我们表明该方法可直接用于全细胞或匀浆生物材料，无需任何提取步骤。所测试的细胞成分以及用于细胞提取的大多数试剂均不干扰该反应。筛选基于使用相对量的比率（脂质/DNA和脂质/蛋白质）来观察生理变化。我们的结果表明，当细胞的磷脂含量发生改变时，可以检测到这些比率的显著差异。所需的样本操作极少且可以自动化。用作工程突变菌株的初步筛选和/或表征时，该测试可能会促使对细胞中脂质的性质和分布进行进一步研究。
