The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.
A rapid method for efficient gene replacement in the filamentous fungus Aspergillus nidulans
Open accessAbstract
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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Citations
Cites
Attenuated virulence of uridine-uracil auxotrophs of Aspergillus fumigatus
D'Enfert, C., Diaquin, M., Delit, A., Wuscher, N., Debeaupuis, J. P., Huerre, M., & Latge, J. P. (1996) · Infection and Immunity · doi:10.1128/iai.64.10.4401-4405.1996 · Cited by 123
Cited by
Bikard, D., Jiang, W., Samai, P., Hochschild, A., Zhang, F., & Marraffini, L. A. (2013) · Nucleic Acids Research · doi:10.1093/nar/gkt520 · Cited by 1,145
da Silva Ferreira, M. E., Kress, M. R. V. Z., Savoldi, M., Goldman, M. H. S., Härtl, A., Heinekamp, T., Brakhage, A. A., & Goldman, G. H. (2006) · Eukaryotic Cell · doi:10.1128/ec.5.1.207-211.2006 · Cited by 403
Scheckhuber, C. Q., Erjavec, N., Tinazli, A., Hamann, A., Nyström, T., & Osiewacz, H. D. (2007) · Nature Cell Biology · doi:10.1038/ncb1524 · Cited by 344
Ferrières, L., Hémery, G., Nham, T., Guérout, A.-M., Mazel, D., Beloin, C., & Ghigo, J.-M. (2010) · Journal of Bacteriology · doi:10.1128/jb.00621-10 · Cited by 339
λ Red-Mediated Genetic Manipulation of Antibiotic-Producing Streptomyces
Gust, B., Chandra, G., Jakimowicz, D., Yuqing, T., Bruton, C. J., & Chater, K. F. (2004) · Advances in Applied Microbiology · doi:10.1016/s0065-2164(04)54004-2 · Cited by 272
Metadata
- Authors
- Chaveroche, M.-K.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2000-11-15
- Volume
- 28
- Issue
- 22
- Pages
- 97e-97
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- OpenAlex
- W2111198831
- Cited by
- 428
- References
- 1
- FWCI
- 3.15
- Type
- journal article
- Language
- en
- Open access
- bronze
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- Chaveroche, M.-K. (2000). A rapid method for efficient gene replacement in the filamentous fungus Aspergillus nidulans. Nucleic Acids Research. https://doi.org/10.1093/nar/28.22.e97
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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