Addition of exogenous polypeptides on the mammalian reovirus outer capsid using reverse genetics

J Virol Methods. 2012 Feb;179(2):342-50. doi: 10.1016/j.jviromet.2011.11.021. Epub 2011 Dec 1.

Abstract

Addition of exogenous peptide sequences on viral capsids is a powerful approach to study the process of viral infection or to retarget viruses toward defined cell types. Until recently, it was not possible to manipulate the genome of mammalian reovirus and this was an obstacle to the addition of exogenous sequence tags onto the capsid of a replicating virus. This obstacle has now been overcome by the availability of the plasmid-based reverse genetics system. In the present study, reverse genetics was used to introduce different exogenous peptides, up to 40 amino acids long, at the carboxyl-terminal end of the σ1 outer capsid protein. The tagged viruses obtained were infectious, produce plaques of similar size, and could be easily propagated at high titers. However, attempts to introduce a 750 nucleotides-long sequence failed, even when it was added after the stop codon, suggesting a possible size limitation at the nucleic acid level.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Capsid Proteins / genetics*
  • Cell Line
  • Chlorocebus aethiops
  • Gene Expression
  • Genetic Engineering / methods
  • Genetic Vectors
  • Mice
  • Orthoreovirus, Mammalian / genetics*
  • Plasmids
  • Recombinant Fusion Proteins / genetics*
  • Reverse Genetics / methods*

Substances

  • Capsid Proteins
  • Recombinant Fusion Proteins