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使用盐酸胍从RNA肿瘤病毒中分离RNA和蛋白质。

The use of guanidine-HCl for the isolation of both RNA and protein from RNA tumour viruses.

作者信息

Pavlovec A, Evenson D P, Hamilton M G

出版信息

J Gen Virol. 1978 Jul;40(1):239-43. doi: 10.1099/0022-1317-40-1-239.

Abstract

The RNA components of two C-type RNA viruses, avian myeloblastosis virus and Friend leukaemia virus, have been isolated by treatment of the viruses with 6 M-guanidine-HCl and precipitation with ethanol. The virus proteins were recovered by lyophilization of the guanidine-HCl-ethanol supernatant after thorough dialysis against 0.5 mM-dithiothreitol. This simple method yielded RNA of similar quality to the phenol and sodium dodecyl sulphate (SDS) extraction methods, and the same amount of 60-70S RNA, although a fraction of the smaller (4S) species remained in the protein fraction. The sedimentation patterns of heat-denatured RNA extracted by either method were similar. Electrophoretic analyses of the extracted proteins in polyacrylamide gel gradients containing SDS gave patterns that were very similar to those obtained by direct analysis of SDS disrupted viruses.

摘要

通过用6M盐酸胍处理禽成髓细胞瘤病毒和弗瑞德白血病病毒这两种C型RNA病毒,并使用乙醇沉淀,已分离出它们的RNA成分。在用0.5mM二硫苏糖醇充分透析后,通过对盐酸胍-乙醇上清液进行冻干回收病毒蛋白。这种简单方法产生的RNA质量与苯酚和十二烷基硫酸钠(SDS)提取方法相似,且60 - 70S RNA的量相同,尽管较小的(4S)部分仍留在蛋白质部分中。用这两种方法提取的热变性RNA的沉降模式相似。在含有SDS的聚丙烯酰胺凝胶梯度中对提取的蛋白质进行电泳分析,得到的图谱与直接分析SDS裂解病毒所获得的图谱非常相似。

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