| RRC ID |
31009
|
| Author |
Faix J, Kreppel L, Shaulsky G, Schleicher M, Kimmel AR.
|
| Title |
A rapid and efficient method to generate multiple gene disruptions in Dictyostelium discoideum using a single selectable marker and the Cre-loxP system.
|
| Journal |
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.
|
| Volume |
32(19)
|
| Pages |
e143
|
| Published |
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
|
| Times Cited |
164
|
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| Cellular slime molds |
G90002
G90003 |
