ArticleNucleic acids research2000
A rapid method for efficient gene replacement in the filamentous fungus Aspergillus nidulans.
Article in Nucleic acids research, 2000. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 222 papers.
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222 citing papers in PubMed.
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- Deciphering the Causes of IbfA-Mediated Abortive Infection in the P22-like Phage UAB_Phi20.International journal of molecular sciences · 2025Article
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- Considerations for Domestication of Novel Strains of Filamentous Fungi.ACS synthetic biology · 2025Review
- Combined inactivation of the SOS response with TCA fumarases and the adaptive response enhances antibiotic susceptibility againstFrontiers in microbiology · 2025Article
- RES-Xre toxin-antitoxin locusEmerging microbes & infections · 2024Article
- Aspergillus fumigatus conidial surface-associated proteome reveals factors for fungal evasion and host immunity modulation.Nature microbiology · 2024Article
- CRISPR/Cas9 mediated targeted knock-in of eglA gene to improve endoglucanase activity of Aspergillus fumigatus LMB-35Aa.Scientific reports · 2024Article
- Carbapenem-resistant Klebsiella pneumoniae capsular types, antibiotic resistance and virulence factors in China: a longitudinal, multi-centre study.Nature microbiology · 2024Article
- Aspergillus fumigatus mitogen-activated protein kinase MpkA is involved in gliotoxin production and self-protection.Nature communications · 2024Article
- No fitness cost entailed by type VI secretion system synthesis, assembly, contraction, or disassembly in enteroaggregativeJournal of bacteriology · 2023Article
162 more citing papers are in PubMed but not listed here.
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3 authors.
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Abstract
The construction of mutant fungal strains is often limited by the poor efficiency of homologous recombination in these organisms. Higher recombination efficiencies can be obtained by increasing the length of homologous DNA flanking the transformation marker, although this is a tedious process when standard molecular biology techniques are used for the construction of gene replacement cassettes. Here, we present a two-step technology which takes advantage of an Escherichia coli strain expressing the phage lambda Red(gam, bet, exo) functions and involves (i) the construction in this strain of a recombinant cosmid by in vivo recombination between a cosmid carrying a genomic region of interest and a PCR-generated transformation marker flanked by 50 bp regions of homology with the target DNA and (ii) genetic exchange in the fungus itself between the chromosomal locus and the circular or linearized recombinant cosmid. This strategy enables the rapid establishment of mutant strains carrying gene knock-outs with efficiencies >50%. It should also be appropriate for the construction of fungal strains with gene fusions or promoter replacements.
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