Abstract
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 cell-free 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.
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