Efficient PCR-based gene targeting with a recyclable marker for Aspergillus nidulans.
Fungal Genet Biol
; 43(1): 54-64, 2006 Jan.
Article
em En
| MEDLINE
| ID: mdl-16289954
ABSTRACT
The rapid accumulation of genomic sequences from a large number of eukaryotes, including numerous filamentous fungi, has created a tremendous scientific potential, which can only be realized if precise site-directed genome modifications, like gene deletions, promoter replacements, in-frame GFP fusions and specific point mutations can be made rapidly and reliably. The development of gene-targeting techniques in filamentous fungi and other higher eukaryotes has been hampered because foreign DNA is predominantly integrated randomly into the genome. For Aspergillus nidulans, we have developed a flexible method for gene-targeting employing a bipartite gene-targeting substrate. This substrate is made solely by PCR, which obviates the need for bacterial subcloning steps. The method reduces the number of false positives and can be used to produce virtually any genome alteration. A major advance of the method is that it allows multiple subsequent genome manipulations to be performed as the selectable marker is recycled.
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Base de dados:
MEDLINE
Assunto principal:
Aspergillus nidulans
/
Marcação de Genes
/
Genes Fúngicos
Idioma:
En
Revista:
Fungal Genet Biol
Assunto da revista:
GENETICA
/
MICROBIOLOGIA
Ano de publicação:
2006
Tipo de documento:
Article
País de afiliação:
Dinamarca