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Formation of bacteriophage MS2 infectious units in a cell‐free translation system
Author(s) -
L. Katanaev Vladimir,
S. Spirin Alexander,
Reuss Matthias,
Siemann Martin
Publication year - 1996
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(96)01158-1
Subject(s) - bacteriophage ms2 , bacteriophage , translation (biology) , virology , chemistry , translation system , biology , biochemistry , escherichia coli , gene , messenger rna , in vitro
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.