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Gene. 2002 Aug 21;296(1-2):179-85.

Construction of a mariner-based transposon for epitope-tagging and genomic targeting.

Author information

1
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA.

Abstract

Mobile genetic elements are often employed for constructing gene fusions or to perform mutagenesis. mariner transposons are well-suited to such applications because of their low site specificity, in vitro activity, and exceptionally broad host range. This report describes a mariner-based method for rapidly creating a large number of insertion mutants that can be converted to in-frame epitope fusions in a single step. First, a mariner-based vector is used to deliver a FLP recombinase substrate randomly into a target molecule. Expression of the FLP recombinase is then induced to catalyse the excision of sequences flanked by FLP recombinase target recognition sites, leaving behind a triple-FLAG epitope. The reversibility of the excision event provides opportunities for using genomic targeting methods easily to create transcriptional or translational fusions to genes of interest.

PMID:
12383515
[Indexed for MEDLINE]

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