TnFLXopen: Markerless Transposons for Functional Fluorescent Fusion Proteins and Protein Interaction Prediction.

ABSTRACT: Fluorescence microscopy of cells expressing proteins translationally linked to a fluorophore can be a powerful tool to investigate protein localization dynamics in vivo. One major obstacle to reliably analyze biologically relevant localization is the construction of a fusion protein that is both fluorescent and functional. Here, we develop a strategy to construct fluorescent fusions at theoretically any location in the protein by using TnFLXopen random transposon mutagenesis to randomly insert a gene encoding a fluorescent protein. Moreover, insertions within a target gene are enriched by an inducible gene-trap strategy and selection by fluorescence activated cell sorting. Using this approach, we isolate a variety of fluorescent fusions to FtsZ that exhibit ring-like localiza

SUBMITTER: Dempwolff F 

PROVIDER: S-EPMC9241775 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature