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[脑心肌炎病毒双链RNA的熔解与重缔合]

[Melting and reassociation of double-stranded RNA of the encephalomyocarditis virus].

作者信息

Chumakov K M

出版信息

Biokhimiia. 1979 Jan;44(1):57-66.

PMID:217455
Abstract

The processes of melting and reassociation of double-stranded RNA in dimethylsulfoxide were studied. The addition of a small amount of LiCl results in great results in great reduction of Tm (temperature of melting), whereas the NaCl produces the opposite effect. It is suggested, that LiCl coordinates the molecules of H2O, reducing their activity, and consequently destabilises dsRNA. Mild conditions for melting and reassociation of RNA can be created. It was found that under optimal conditions for dsRNA melting, the degree of strand separation depends on the overall concentration of RNA, irrespective of the type of RNA added to the dsRNA preparation. Reassociation of dsRNA of EMC virus proceeds much faster than that of dsRNA of a related poliovirus. Addition of poly(C) to an annealing mixture slows down the rate of reassociation of EMC dsRNA, producing no effect on the poliovirus dsRNA reassociation. It is suggested that the presence of large poly(C) and poly(G) tracts in the complementary strands of the RNA determines its anomalous fast reassociation. Upon incubation of completely separated strands of EMC dsRNA in a water solution with high ionic strength partially double-stranded aggregates are formed. The formation of aggregates is prevented by addition of poly(A), which indicates that they are produced by "zippening" of a molecule starting with poly(A):poly(U) region. The significance of homopolymeric regions for stability of dsRNA of the EMC virus as well as their role in viral multiplication are discussed.

摘要

研究了双链RNA在二甲基亚砜中的熔化和重新缔合过程。添加少量LiCl会导致熔解温度(Tm)大幅降低,而NaCl则产生相反的效果。有人认为,LiCl使水分子配位,降低其活性,从而使双链RNA不稳定。可以创造温和的RNA熔化和重新缔合条件。发现在双链RNA熔化的最佳条件下,链分离程度取决于RNA的总浓度,而与添加到双链RNA制剂中的RNA类型无关。脑心肌炎病毒双链RNA的重新缔合比相关脊髓灰质炎病毒的双链RNA快得多。向退火混合物中添加聚(C)会减慢脑心肌炎病毒双链RNA的重新缔合速率,而对脊髓灰质炎病毒双链RNA的重新缔合没有影响。有人认为,RNA互补链中大量聚(C)和聚(G)片段的存在决定了其异常快速的重新缔合。将脑心肌炎病毒双链RNA完全分离的链在高离子强度的水溶液中孵育时,会形成部分双链聚集体。添加聚(A)可防止聚集体的形成,这表明它们是由从聚(A):聚(U)区域开始的分子“拉链”产生的。讨论了同聚物区域对脑心肌炎病毒双链RNA稳定性的意义及其在病毒增殖中的作用。

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