This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sugui, J. A. Articles by Kwon-Chung, K. J. Search for Related Content PubMed PubMed Citation Articles by Sugui, J. A. Articles by Kwon-Chung, K. J. Agricola Articles by Sugui, J. A. Articles by Kwon-Chung, K. J.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, April 2005, p. 1798-1802, Vol. 71, No. 4
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.4.1798-1802.2005

Agrobacterium tumefaciens-Mediated Transformation of Aspergillus fumigatus: an Efficient Tool for Insertional Mutagenesis and Targeted Gene Disruption

Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland

Received 1 September 2004/ Accepted 27 October 2004

Agrobacterium tumefaciens was used to transform Aspergillus fumigatus by either random or site-directed integration of transforming DNA (T-DNA). Random mutagenesis via Agrobacterium tumefaciens-mediated transformation (ATMT) was accomplished with T-DNA containing a hygromycin resistance cassette. Cocultivation of A. fumigatus conidia and Agrobacterium (1:10 ratio) for 48 h at 24°C resulted in high frequencies of transformation (>100 transformants/10⁷ conidia). The majority of transformants harbored a randomly integrated single copy of T-DNA and were mitotically stable. We chose alb1, a polyketide synthase gene, as the target gene for homologous integration because of the clear phenotype difference between the white colonies of alb1 mutant strains and the bluish-green colonies of wild-type strains. ATMT with a T-DNA-containing alb1 disruption construct resulted in 66% albino transformants. Southern analysis revealed that 19 of the 20 randomly chosen albino transformants (95%) were disrupted by homologous recombination. These results suggest that ATMT is an efficient tool for transformation, random insertional mutagenesis, and gene disruption in A. fumigatus.

This article has been cited by other articles:

• Ma, Y., Qiao, J., Liu, W., Wan, Z., Wang, X., Calderone, R., Li, R. (2008). The Sho1 Sensor Regulates Growth, Morphology, and Oxidant Adaptation in Aspergillus fumigatus but Is Not Essential for Development of Invasive Pulmonary Aspergillosis. Infect. Immun. 76: 1695-1701 [Abstract] [Full Text]  
• Smulian, A. G., Gibbons, R. S., Demland, J. A., Spaulding, D. T., Deepe, G. S. Jr. (2007). Expression of Hygromycin Phosphotransferase Alters Virulence of Histoplasma capsulatum. Eukaryot Cell 6: 2066-2071 [Abstract] [Full Text]  
• Sugui, J. A., Pardo, J., Chang, Y. C., Zarember, K. A., Nardone, G., Galvez, E. M., Mullbacher, A., Gallin, J. I., Simon, M. M., Kwon-Chung, K. J. (2007). Gliotoxin Is a Virulence Factor of Aspergillus fumigatus: gliP Deletion Attenuates Virulence in Mice Immunosuppressed with Hydrocortisone. Eukaryot Cell 6: 1562-1569 [Abstract] [Full Text]  
• Sugui, J. A., Pardo, J., Chang, Y. C., Mullbacher, A., Zarember, K. A., Galvez, E. M., Brinster, L., Zerfas, P., Gallin, J. I., Simon, M. M., Kwon-Chung, K. J. (2007). Role of laeA in the Regulation of alb1, gliP, Conidial Morphology, and Virulence in Aspergillus fumigatus. Eukaryot Cell 6: 1552-1561 [Abstract] [Full Text]