Converting an FRT-Tagged Gene into a Fluorescent Protein Gene Fusion by Flp-Mediated Site-Specific Recombination.

Recombination, Genetic DNA Nucleotidyltransferases / genetics Plasmids / genetics Gene Fusion Genes, Reporter

Journal

Cold Spring Harbor protocols
ISSN: 1559-6095
Titre abrégé: Cold Spring Harb Protoc
Pays: United States
ID NLM: 101524530

Informations de publication

Date de publication:
01 09 2023
Historique:
medline: 4 9 2023
pubmed: 23 2 2023
entrez: 22 2 2023
Statut: epublish

Résumé

This protocol uses conditional plasmids carrying the open reading frame (orf) of either superfolder green fluorescent protein (sfGFP) or monomeric Cherry (mCherry) fused to a flippase (Flp) recognition target (FRT) site. In cells expressing the Flp enzyme, site-specific recombination between the plasmid-borne FRT and an FRT "scar" in a target gene in the bacterial chromosome results in chromosomal integration of the plasmid with the concomitant in-frame fusion of the target gene to the fluorescent protein orf. This event can be positively selected using an antibiotic-resistance marker (

Identifiants

DOI: 10.1101/pdb.prot107951 PMID: 36813484
pubmed: 36813484
pii: pdb.prot107951
doi: 10.1101/pdb.prot107951
doi:

Substances chimiques

DNA Nucleotidyltransferases EC 2.7.7.-

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

663-670

Informations de copyright

© 2023 Cold Spring Harbor Laboratory Press.

Auteurs

Roberto Balbontín (R)

Departamento de Genética, Facultad de Biología, Universidad de Sevilla, 41080 Sevilla, Spain.

Mathilde Ratel (M)

Université Paris-Saclay, CEA, CNRS, Institut de Biologie Intégrative de la Cellule (I2BC), 91190 Gif-sur-Yvette, France.

Nara Figueroa-Bossi (N)

Université Paris-Saclay, CEA, CNRS, Institut de Biologie Intégrative de la Cellule (I2BC), 91190 Gif-sur-Yvette, France.
ORCID: 0000-0002-7552-0225

Lionello Bossi (L)

Université Paris-Saclay, CEA, CNRS, Institut de Biologie Intégrative de la Cellule (I2BC), 91190 Gif-sur-Yvette, France lionello.bossi@i2bc.paris-saclay.fr.

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Classifications MeSH

questionsmedicales.fr › Phénomènes génétiques › Recombinaison génétique › Converting an FRT-Tagged Gene into a...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Transferases › Phosphotransferases › Nucleotidyltransferases › DNA nucleotidyltransferases › Converting an FRT-Tagged Gene into a...
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Plasmides › Converting an FRT-Tagged Gene into a...
questionsmedicales.fr › Phénomènes génétiques › Recombinaison génétique › Fusion de gènes › Converting an FRT-Tagged Gene into a...
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Génome › Composants de génome › Gènes › Gènes rapporteurs › Converting an FRT-Tagged Gene into a...