HUFF E, OXLEY H, SILVERMAN C S
J Bacteriol. 1964 Oct;88(4):1155-62. doi: 10.1128/jb.88.4.1155-1162.1964.
Huff, Eskin (National Institute of Allergy and Infectious Diseases, Bethesda, Md.), Harriet Oxley, and Carol S. Silverman. Density-gradient patterns of Staphylococcus aureus cells and cell walls during growth and mechanical disruption. J. Bacteriol. 88:1155-1162. 1964.-Procedures capable of rapid disruption of Staphylococcus aureus cells with optimal release of intact cell walls were investigated. This search was implemented by observation of the flotation patterns of cells and subcellular particulate matter after centrifugation in a cesium chloride density gradient. A quantitative evaluation of the light-scatter throughtout the gradient was achieved by transfer of the entire density gradient into an optical cell with wedge-shaped cross section. When this cell was photographed under indirect illumination, each band of light-scattering material appeared on the negative as a shaded curve, with an area proportional to amount of that material present. A series of photographs of known amounts of cells and cell walls was used to estimate the amounts of these materials in mixtures of the two occurring during mechanical disruption. With the methods employed, time studies established the optimal time for release of cell walls as 5 min in a Braun shaker. The use of sucrose gradients in the further purification of cell walls, and chemical analysis of the isolated walls, are described.
赫夫、埃斯金（美国国立过敏与传染病研究所，马里兰州贝塞斯达）、哈丽雅特·奥克斯利和卡罗尔·S·西尔弗曼。金黄色葡萄球菌细胞及细胞壁在生长和机械破碎过程中的密度梯度模式。《细菌学杂志》88:1155 - 1162。1964年。——研究了能够快速破碎金黄色葡萄球菌细胞并使完整细胞壁最佳释放的方法。通过观察在氯化铯密度梯度中离心后细胞和亚细胞颗粒物质的漂浮模式来进行此项探索。通过将整个密度梯度转移到具有楔形横截面的光学池中，实现了对整个梯度中光散射的定量评估。当在间接照明下拍摄这个池子时，每条光散射物质带在底片上呈现为一条阴影曲线，其面积与该物质的含量成正比。使用一系列已知量的细胞和细胞壁的照片来估计在机械破碎过程中这两者混合物中这些物质的量。采用这些方法进行的时间研究确定了在 Braun 振荡器中细胞壁释放的最佳时间为5分钟。描述了在细胞壁进一步纯化中使用蔗糖梯度以及对分离出的细胞壁进行化学分析的情况。
