期刊文献

Formation of bacteriophage MS2 infectious units in a cell-free translation system 收藏

在无细胞翻译系统中形成噬菌体MS2感染单位

原文求助发布源

作者

Vladimir L. Katanaev[1,2,1]; Alexander S. Spirin[2]; Matthias Reuss[1] and Martin Siemann[1];

作者单位

1 Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany 2 Institute of Protein Research, Pushchino, Moscow Region 142292, Russian Federation

关键词

Cell-free translation; In vitro phage assembly; RNA-protein interaction; Infectivity; Phage MS2 morphogenesis; RNA vector delivery system; Molecular evolution; PAGE, polyacrylamide gel electrophoresis; PFU, plaque forming unit

关键词译文

无细胞翻译;体外噬菌体组装; RNA蛋白相互作用;感染;噬菌体MS2形态发生; RNA载体递送系统;分子进化; PAGE，聚丙烯酰胺凝胶电泳; PFU，斑块形成单位

页码

143–148

DOI

10.1016/S0014-5793(96)01158-1

来源信息

FEBS Letters ISSN：e1873-3468, 1996年, 397卷, 2-3期, 143–148页

摘要

We show that a simple cell-free translation system from Escherichia coli, programmed with phage MS2 RNA, is able to infect F+ E. coli cells. The plaques appearing on the E. coli host strain are morphologically indistinguishable from those derived from normal phage MS2 infection. This effect is strictly translation-dependent, since an incomplete translation system or the system inhibited by antibiotics leads to no infection. The cellfree based infection is maximal under conditions favouring the highest synthesis of maturation protein (one of the four phage-encoded proteins). The infection is abolished when RNase A or trypsin treatment is included before addition of cells. Similarly, due to RNA and maturation protein degradation, the continued incubation of the translation mixture under protein synthesis conditions significantly decreases infectivity. These findings suggest the formation of ‘minimal infectious units’, simple complexes of MS2 RNA and maturation protein. Here we describe the first example of bacteriophage infectious unit formation directly performed in a cell-free translation system. A possible application of this phenomenon might be the construction of newly designed RNA vector delivery systems and, moreover, could be an approach for molecular evolution studies. .overlined { text-decoration: overline; } .struck { text-decoration:line-through; } .underlined { text-decoration:underline; } .doubleUnderlined { text-decoration:underline;border-bottom:1px solid #000; } Enhanced Article (HTML) Get PDF (677K)Get PDF (677K) More content like thisFind more content: like this articleFind more content written by: Vladimir L. Katanaev Alexander S. Spirin Matthias Reuss Martin Siemann All Authors

摘要译文

我们显示，使用噬菌体MS2 RNA编程的大肠杆菌的简单无细胞翻译系统能够感染F +E coli细胞。Eques上出现的斑块大肠杆菌宿主菌在形态学上与来自正常噬菌体MS2感染的菌株形态学上无区别。这种作用是严格的依赖于翻译，由于不完整的翻译系统或被抗生素抑制的系统导致没有感染在有利于成熟蛋白质的最高合成（四种噬菌体编码的蛋白质之一）的条件下，基于无细胞的感染是最大的在添加细胞之前，包括RNase A或胰蛋白酶处理时，感染被消除。同样地，由于RNA和成熟蛋白质降解，翻译混合物在蛋白质合成条件下的继续孵育显着降低了感染性。这些发现表明形成了最小的感染单位MS2 RNA和成熟蛋白的简单复合物我们描述了在无细胞翻译系统中直接进行的噬菌体感染单位形成的第一个例子这种现象的可能应用可能是新设计的RNA载体递送系统的构建，此外，可能是分子进化研究的一种方法重叠的{text-decoration：overline;竤卡车{text-decoration：line-through; }竨nderlined {text-decoration：underline; }竏oubleUnderlined {text-decoration：underline; border-bottom：1px solid＃000;}增强文章（HTML）获取PDF（677K）获取PDF（677K）更多内容喜欢此文章更多内容：像这篇文章查找更多内容：VladimirL KatanaevAlexanderS Spirin Matthias Reuss Martin Siemann所有作者

Vladimir L. Katanaev[1,2,1]; Alexander S. Spirin[2]; Matthias Reuss[1] and Martin Siemann[1];. Formation of bacteriophage MS2 infectious units in a cell-free translation system[J]. FEBS Letters, 1996,397(2-3): 143–148