Construct conversions caused by simultaneous co-transfection: "GFP-walking"

  • Bestvater F
  • Knoch T
  • Langowski J
 et al. 
  • 8


    Mendeley users who have this article in their library.
  • 9


    Citations of this article.


Several GFP variants have been developedfor multicolor labeling in vivo. Here we report that simultaneous co-transfection of fluorescent protein chimeras can give false-positive results caused by the conversion of spectral properties. Under standard transfection conditions, approximately 8% of cells produce false-positive results, but, depending on the conditions, up to 26% of the cells permanently express altered fusion proteins. This compromises the interpretation of the results. The conversion is independent of transfection methods or cell types. Our results show that the effect is based on homologous recombination/repair/replication process events that occur between the nucleotide sequences of the fluorescent proteins. Consecutive transfection or low sequence similarities avoided recombination. The appearance of conversion facilitates exchanges of spectral properties infusion proteins, the creation of libraries, or the assembly of DNA fusion constructs in vivo. The detailed quantification of the conversion rate allows the investigation of recombination/repair/replication processes in general.

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

  • SCOPUS: 2-s2.0-0036213945
  • ISSN: 07366205
  • SGR: 0036213945
  • PMID: 11962606
  • PUI: 34298712
  • ISBN: 07366205


  • Felix Bestvater

  • Tobias A. Knoch

  • Jörg Langowski

  • Eberhard Spiess

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free