KELLENBERGER E, RYTER A, SECHAUD J
J Biophys Biochem Cytol. 1958 Nov 25;4(6):671-8. doi: 10.1083/jcb.4.6.671.
The nucleoids of Escherichia coli, independently of the physiological state of the bacteria, are shown to be preserved as a fine-stranded fibrillar nucleoplasm by an OsO(4) fixation under defined conditions: acetate-veronal buffer pH 6, presence of Ca(++) and amino acids, stabilization with uranyl-acetate before dehydration. The same fixation procedure applied to the DNA of vegetative phage reveals a pool of homogeneous fibrillar structure very similar to the nucleoplasm. The "versene test," which produces a coarse coagulation of these plasms, emphasizes the similar behaviour of the pool and the nucleoids. The heads of mature phage are preserved in their true polyhedral shape by the standard fixation procedure, although they may be badly distorted when fixed under different conditions. Lanthanum nitrate and uranyl-acetate are shown to increase markedly the contrast of both phage and cytoplasm. The consequences of the fibrillar structure of the genetic material are discussed in relation to the probable division process.
在特定条件下(pH 6的醋酸-韦罗那缓冲液、存在Ca(++)和氨基酸、脱水前用醋酸铀酰稳定),通过四氧化锇固定,结果表明,无论细菌的生理状态如何,大肠杆菌的类核都能保存为细链状的纤维状核质。将相同的固定程序应用于营养期噬菌体的DNA,可发现一组与核质非常相似的均匀纤维状结构。“依地酸盐试验”会使这些质体发生粗凝聚,这突出了该组结构与类核的相似行为。通过标准固定程序,成熟噬菌体的头部能保持其真正的多面体形状,不过在不同条件下固定时可能会严重变形。硝酸镧和醋酸铀酰可显著提高噬菌体和细胞质的对比度。结合可能的分裂过程,讨论了遗传物质纤维状结构的影响。