An in vitro DNA virus for in vitro protein evolution

In vitro virus is a molecular construct for in vitro protein evolution, whi ch requires some mechanism to link phenotype to genotype. The first in vitr o virus was realized by bonding a nascent protein with its coding mRNA via puromycin in in vitro translation. We report a new construct of in vitro DN A virus. The virion was a covalent cDNA-protein fusion, and virion formatio n did not require any modification of mRNA. Due to intactness of mRNA, this type of in vitro DNA virus will take the next step toward in vitro autonom ous evolution, just like in vivo viral evolution in a cellstat. (C) 2001 Fe deration of European Biochemical Societies. Published by Elsevier Science B .V. All rights reserved.