RRC ID 31009
著者 Faix J, Kreppel L, Shaulsky G, Schleicher M, Kimmel AR.
タイトル A rapid and efficient method to generate multiple gene disruptions in Dictyostelium discoideum using a single selectable marker and the Cre-loxP system.
ジャーナル Nucleic Acids Res
Abstract Dictyostelium discoideum has proven an exceptionally powerful system for studying numerous aspects of cellular and developmental functions. The relatively small ( approximately 34 Mb) chromosomal genome of Dictyostelium and high efficiency of targeted gene disruption have enabled researchers to characterize many specific gene functions. However, the number of selectable markers in Dictyostelium is restricted, as is the ability to perform effective genetic crosses between strains. Thus, it has been difficult to create multiple mutations within an individual cell to study epistatic relationships among genes or potential redundancies between various pathways. We now describe a robust system for the production of multiple gene mutations in Dictyostelium by recycling a single selectable marker, Blasticidin S resistance, using the Cre-loxP system. We confirm the effectiveness of the system by generating a single cell carrying four separate gene disruptions. Furthermore, the cells remain sensitive to transformation for additional targeted or random mutagenesis requiring Blasticidin selection and for functional expression studies of mutated or tagged proteins using other selectable markers.
巻・号 32(19)
ページ e143
公開日 2004-10-26
DOI 10.1093/nar/gnh136
PII 32/19/e143
PMID 15507682
PMC PMC528815
MeSH Animals Dictyostelium / drug effects Dictyostelium / genetics* Drug Resistance Gene Targeting / methods* Genetic Markers Integrases / metabolism* Mutation Nucleosides / pharmacology Recombination, Genetic Viral Proteins / metabolism*
IF 11.502
引用数 164
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
細胞性粘菌 G90002 G90003