Koenig R, An D, Burgermeister W
Institut für Viruskrankheiten der Pflanzen, Biologischen Bundesanstalt für Land- und Forstwirtschaft, Braunschweig, F.R.G.
J Virol Methods. 1988 Jan;19(1):57-68. doi: 10.1016/0166-0934(88)90007-9.
In attempts to use dot-blot hybridization tests for the identification of viruses or for assigning them to a certain taxonomic group we found that hybridization signals may be given not only by the homologous virus, but also by heterologous viruses belonging to the same or different taxonomic groups. Possible reasons for this phenomenon, which was observed with uncloned as well as with cloned cDNAs, are discussed. Quantitative dot-blot hybridization tests with extracted viral RNAs proved to be very sensitive in differentiating closely related viruses which were barely distinguishable in serological tests. Estimates on the degree of homology between the RNAs of different viruses may be influenced by a number of experimental parameters, such as competition for the available cDNA between homologous and heterologous RNAs or homologous RNAs in different concentrations on the same sheet of nitrocellulose, saturation phenomena due to close packaging of highly concentrated RNA on the blot and, of course, stringency conditions during washing procedures. Taking these parameters into account we have reestimated the degree of homology between the RNAs of 5 tombusviruses. Our new data suggest that the order of sequence for the relationships among these 5 tombusviruses is similar to that proposed by Koenig and Gibbs (1986) on the basis of serological data.
在尝试使用斑点杂交试验来鉴定病毒或将其归入某一分类学类别时，我们发现杂交信号不仅可能由同源病毒产生，也可能由属于相同或不同分类学类别的异源病毒产生。本文讨论了这种现象（在未克隆的以及克隆的cDNA中均观察到）可能的原因。用提取的病毒RNA进行的定量斑点杂交试验在区分血清学试验中几乎无法区分的密切相关病毒方面被证明非常灵敏。不同病毒RNA之间同源性程度的估计可能受到许多实验参数的影响，例如同源和异源RNA之间对可用cDNA的竞争、同一硝酸纤维素膜上不同浓度同源RNA之间的竞争、印迹上高度浓缩RNA紧密堆积导致的饱和现象，当然还有洗涤过程中的严格条件。考虑到这些参数，我们重新估计了5种番茄丛矮病毒RNA之间的同源性程度。我们的新数据表明，这5种番茄丛矮病毒之间关系的序列顺序与柯尼希和吉布斯（1986年）根据血清学数据提出的顺序相似。
