Zuidema D, Bierhuizen M F, Cornelissen B J, Bol J F, Jaspars E M
Virology. 1983 Mar;125(2):361-9. doi: 10.1016/0042-6822(83)90208-8.
The largest genome segment, RNA 1, of alfalfa mosaic virus forms complexes with viral coat protein. These complexes were subjected to digestion with ribonucleases T1 or A and filtered onto Millipore filters. Specific fragments were collected from the filters by phenol extraction. After electrophoretic separation in denaturing polyacrylamide gels, these fragments were sequenced. Besides extracistronic fragments originating from the 3'-terminal region of RNA 1, fragments were found originating from an intracistronic region of the RNA. A striking phenomenon is that the intracistronic fragments were not found when ribonuclease A was used to degrade RNA/protein complexes. The findings are in agreement with the postulation of Houwing and Jaspars (1978), that a conformational change at the 3' ends of the genome RNAs induced by the coat protein is a prerequisite to start an infection cycle.
苜蓿花叶病毒最大的基因组片段RNA 1与病毒外壳蛋白形成复合物。这些复合物用核糖核酸酶T1或A进行消化,然后过滤到微孔滤膜上。通过苯酚抽提从滤膜上收集特定片段。在变性聚丙烯酰胺凝胶中进行电泳分离后,对这些片段进行测序。除了源自RNA 1 3'末端区域的顺反子外片段,还发现了源自该RNA顺反子区域内的片段。一个显著的现象是,当使用核糖核酸酶A降解RNA/蛋白质复合物时,未发现顺反子内片段。这些发现与Houwing和Jaspars(1978年)的假设一致,即外壳蛋白诱导的基因组RNA 3'末端构象变化是启动感染周期的先决条件。