Boyce F M, Sundin O, Barsoum J, Varshavsky A
J Virol. 1982 Apr;42(1):292-6. doi: 10.1128/JVI.42.1.292-296.1982.
A new method for the isolation of simian virus 40 nucleoprotein complexes from nuclei of lytically infected cells is described. The method is based on the addition of a thiol-specific reagent, 5'5'-dithiobis(2-nitrobenzoic acid), to lysis and extraction buffers. By inhibiting an uncoating activity during simian virus 40 extraction, 5'5'-dithiobis (2-nitrobenzoic acid) allows the use of efficient extraction buffers, such as one containing Triton X-100 and EDTA, for the isolation of native simian virus 40 minichromosomes and virion-type structures. Use of the method is illustrated by following encapsidation of simian virus 40 minichromosomes in a pulse-chase experiment. Since 5'5'-dithiobis (2-nitrobenzoic acid) is an inhibitor of many different enzymes, the 5',5'-dithiobis (2-nitrobenzoic acid) extraction technique may be useful for the isolation of not only papovaviruses but also other viruses and possibly cellular chromatin.
本文描述了一种从裂解感染细胞的细胞核中分离猿猴病毒40核蛋白复合物的新方法。该方法基于在裂解和提取缓冲液中添加一种硫醇特异性试剂，即5,5'-二硫代双（2-硝基苯甲酸）。通过在猿猴病毒40提取过程中抑制脱壳活性，5,5'-二硫代双（2-硝基苯甲酸）使得能够使用高效的提取缓冲液，如含有 Triton X-100 和 EDTA 的缓冲液，来分离天然的猿猴病毒40微型染色体和病毒粒子型结构。在脉冲追踪实验中通过追踪猿猴病毒40微型染色体的衣壳化过程来说明该方法的应用。由于5,5'-二硫代双（2-硝基苯甲酸）是许多不同酶的抑制剂，5,5'-二硫代双（2-硝基苯甲酸）提取技术可能不仅对乳头瘤病毒的分离有用，而且对其他病毒以及可能的细胞染色质的分离也有用。
