Wiegand, S. ; Kruse, J. ; Gronemann, S. ; Hammann, Christian (2011)
Efficient generation of gene knockout plasmids for Dictyostelium discoideum using one-step cloning.
In: Genomics, 97 (5)
doi: 10.1016/j.ygeno.2011.02.001
Artikel, Bibliographie
Kurzbeschreibung (Abstract)
The amoeba Dictyostelium discoideum is a well-established model organism for studying numerous aspects of cellular and developmental functions. Its rather small (~34Mb) chromosomal genome and the high efficiency of gene disruption by homologous recombination have enabled researchers to dissect various specific gene functions. We describe here the use of one-step cloning for the fast and efficient generation of deletion vectors that are produced in a one-step reaction by inserting two PCR products into an organism-specific, generic acceptor system. This worked efficiently for all 16 tested constructs directed against genes in the amoeba Dictyostelium discoideum. Saving cost and time, the used protocol represents a significant advancement in the generation of such plasmids compared to the conventionally applied restriction enzyme/ligation approach. Using appropriate selection markers, similar systems could also be useful in other organisms, where genes can be knocked out by homologous recombination.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2011 |
| Autor(en): | Wiegand, S. ; Kruse, J. ; Gronemann, S. ; Hammann, Christian |
| Art des Eintrags: | Bibliographie |
| Titel: | Efficient generation of gene knockout plasmids for Dictyostelium discoideum using one-step cloning. |
| Sprache: | Englisch |
| Publikationsjahr: | 2011 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Genomics |
| Jahrgang/Volume einer Zeitschrift: | 97 |
| (Heft-)Nummer: | 5 |
| DOI: | 10.1016/j.ygeno.2011.02.001 |
| Kurzbeschreibung (Abstract): | The amoeba Dictyostelium discoideum is a well-established model organism for studying numerous aspects of cellular and developmental functions. Its rather small (~34Mb) chromosomal genome and the high efficiency of gene disruption by homologous recombination have enabled researchers to dissect various specific gene functions. We describe here the use of one-step cloning for the fast and efficient generation of deletion vectors that are produced in a one-step reaction by inserting two PCR products into an organism-specific, generic acceptor system. This worked efficiently for all 16 tested constructs directed against genes in the amoeba Dictyostelium discoideum. Saving cost and time, the used protocol represents a significant advancement in the generation of such plasmids compared to the conventionally applied restriction enzyme/ligation approach. Using appropriate selection markers, similar systems could also be useful in other organisms, where genes can be knocked out by homologous recombination. |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie > Regulatorische RNAs und Ribozyme ?? fb10_mikrobiologie ?? 10 Fachbereich Biologie |
| Hinterlegungsdatum: | 23 Feb 2011 14:51 |
| Letzte Änderung: | 05 Mär 2013 09:46 |
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