噬菌体T4基因49的功能。I. 极快速沉降DNA的分离及生化特性分析
Function of gene 49 of bacteriophage T4. I. Isolation and biochemical characterization of very fast-sedimenting DNA.
作者信息
Kemper B, Janz E
出版信息
J Virol. 1976 Jun;18(3):992-9. doi: 10.1128/JVI.18.3.992-999.1976.
Abstract
Very fast-sedimenting DNA was isolated from cells after infection with gene 49 defective phage T4. This DNA appeared membrane bound throughout the time after infection and could be isolated either in the membrane-bound form (M-DNA) or free of membrane (released DNA) depending on the lysis procedure. Released DNA formed complexes of marked stability with sedimentation velocities between 1,400S and 2,100S. These complexes did not seem to contain material other than DNA. This was concluded from the results of RNA, protein, and membrane labeling experiments and density analysis. In addition, these complexes were resistant against treatment with n-butanol, phenol. chloroform-methanol, sodium dodecyl sulfate, Sarkosyl, Pronase, RNase, or lysozyme. The observation that more then 90% of the purified very fast-sedimenting DNA is retrapped by magnesium-Sarkosyl crystals (M-band) suggests that the M-band technique may not be sufficient as a test for DNA-membrane attachment.
摘要
用基因49缺陷型噬菌体T4感染细胞后，从中分离出了沉降速度极快的DNA。这种DNA在感染后的整个时间段内似乎都与膜结合，根据裂解程序，它既可以以膜结合形式（M-DNA）分离，也可以无膜分离（释放的DNA）。释放的DNA形成了具有显著稳定性的复合物，沉降速度在1400S至2100S之间。这些复合物似乎只含有DNA，这是通过RNA、蛋白质和膜标记实验以及密度分析的结果得出的结论。此外，这些复合物对正丁醇、苯酚、氯仿 - 甲醇、十二烷基硫酸钠、 Sarkosyl、链霉蛋白酶、核糖核酸酶或溶菌酶的处理具有抗性。超过90%的纯化后沉降速度极快的DNA被镁 - Sarkosyl晶体重新捕获（M带），这一观察结果表明，M带技术作为DNA与膜附着的检测方法可能并不充分。
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