Laemmli U K
Proc Natl Acad Sci U S A. 1975 Nov;72(11):4288-92. doi: 10.1073/pnas.72.11.4288.
High-molecular-weight DNA is known to collapse into very compact particles in a salt solution containing polymers like poly(ethylene oxide) [(EO)n] or polyacrylate. The biological relevance of this phenomenon is suggested by our recent finding that high concentrations of the highly acidic internal peptides found in the mature T4 bacteriophage head, as well as poly(glutamic acid) and poly(aspartic acid), can collapse DNA in a similar manner. The structure of DNAs collapsed by various methods has been studied with electron microscope. We find (EO)n collapses T4 or T7 bacteriophage DNA into compact particles only slightly larger than the size of the T4 and T7 head, respectively. In contrast, polylysine collapses DNA into different types of structures. Double-stranded DNA collapsed with (EO)n is cut by the single-strand specific Neurospora crassa endonuclease (EC 3.1.4.21) into small fragments. Extensive digestion only occurs above the critical concentration of polymer required for DNA collapse, demonstrating the (EO)n-collapsed DNA contains enzyme-vulnerable regions (probably at each fold), which are preferentially attacked. The size of the DNA fragments produced by limit-digestion with the nuclease ranges between 200 and 400 base pairs when DNA is collapsed by (EO)n. Only fragments of DNA which are larger than 600 base pairs are cut by the endonuclease in (EO)n-containing solution.
众所周知,在含有聚环氧乙烷[(EO)n]或聚丙烯酸酯等聚合物的盐溶液中,高分子量DNA会折叠成非常致密的颗粒。我们最近发现,成熟T4噬菌体头部发现的高浓度高酸性内部肽以及聚谷氨酸和聚天冬氨酸,也能以类似方式使DNA折叠,这表明了这种现象的生物学相关性。用电子显微镜研究了通过各种方法折叠的DNA的结构。我们发现(EO)n分别将T4或T7噬菌体DNA折叠成仅略大于T4和T7头部大小的致密颗粒。相比之下,聚赖氨酸将DNA折叠成不同类型的结构。用(EO)n折叠的双链DNA被单链特异性粗糙脉孢菌内切核酸酶(EC 3.1.4.21)切割成小片段。广泛的消化仅在DNA折叠所需的聚合物临界浓度以上发生,这表明(EO)n折叠的DNA含有易受酶攻击的区域(可能在每个折叠处),这些区域优先受到攻击。当DNA被(EO)n折叠时,用核酸酶进行限量消化产生的DNA片段大小在200至400个碱基对之间。在含(EO)n的溶液中,只有大于600个碱基对的DNA片段被内切核酸酶切割。