Targeted disruption of the TGA3 locus in Arabidopsis thaliana.
Plant J
; 7(2): 359-65, 1995 Feb.
Article
en En
| MEDLINE
| ID: mdl-7704051
ABSTRACT
A major drawback to study gene functions in plant systems is the lack of an effective gene knockout strategy. With a large number of plant genes isolated and the accelerating pace by which this collection is growing, the need for their functional analyses at the whole plant level has become increasingly urgent. Here evidence is reported for the first successful disruption of a non-selectable gene in Arabidopsis thaliana by creating a mutant of the TGA3 locus via targeted insertion of the bacterial neo gene conferring kanamycin (Km) resistance. A beta-glucuronidase (GUS) expression unit outside the region of homology was used as a screenable marker to distinguish homologous recombination events from those of ectopic insertions. PCR amplification coupled with Southern blot screening identified two putative homologous recombination events among 2580 Kmr calli. One callus line was subsequently isolated and the structure of the targeted TGA3 allele confirmed by Southern blot analyses. This study demonstrates the feasibility of targeting a non-selectable locus in Arabidopsis. Combined with future improvements in negative selection strategies and efficient transformation methodologies, gene replacement studies in plants could become a routine technique.
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Bases de datos:
MEDLINE
Asunto principal:
Factores de Transcripción
/
Genes de Plantas
/
Arabidopsis
/
Proteínas de Arabidopsis
/
Proteínas de Unión al ADN
Tipo de estudio:
Prognostic_studies
Idioma:
En
Revista:
Plant J
Asunto de la revista:
BIOLOGIA MOLECULAR
/
BOTANICA
Año:
1995
Tipo del documento:
Article