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

FEBS Letters

L. Katanaev, Vladimir¹  Reuss, Matthias¹  Siemann, Martin¹  S. Spirin, Alexander² 

[1] Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany;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;   

DOI  :  10.1016/S0014-5793(96)01158-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 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.

【 授权许可】

Unknown