Journal article
Targeted gene deletion in Aspergillus fumigatus using microbial machinery and a recyclable marker
Journal of microbiological methods, Vol.95(3), pp.373-378
12/2013
DOI: 10.1016/j.mimet.2013.09.021
PMID: 24161898
Abstract
The emerging invasive fungal pathogen Aspergillus fumigatus causes very serious infections among immunocompromised patient populations. While the genome of this pathogen has been sequenced, a major barrier to better understanding the complex biology of this eukaryotic organism is a lack of tools for efficient genetic manipulation. To improve upon this, we have generated a new gene deletion system for A. fumigatus using yeast recombinational cloning and Agrobacterium tumefaciens mediated transformation (ATMT) employing a recyclable marker system. This system reduced the time for generating a gene deletion strain in our hands by two-thirds (12weeks to 3weeks) using minimal human labor, and we demonstrate that it can be used to efficiently generate multiple gene deletions within a single strain.
•Aspergillus fumigatus, a major fungal pathogen, is difficult to transform.•A system was assembled to more easily transform A. fumigatus.•The system utilizes yeast cloning, Agrobacterium transformation and a recyclable marker.•This uses ‘microbial labor’ and allows for the generation of multiple gene deletions.
Details
- Title: Subtitle
- Targeted gene deletion in Aspergillus fumigatus using microbial machinery and a recyclable marker
- Creators
- Joshua B Kieler - Department of Pathology, University of Iowa Carver College of Medicine, 200 Hawkins Drive, Iowa City, IA 52242, United StatesKhanh L Duong - Department of Pathology, University of Iowa Carver College of Medicine, 200 Hawkins Drive, Iowa City, IA 52242, United StatesW. Scott Moye-Rowley - Department of Molecular Physiology and Biophysics, 6-530 Bowen Science Building, University of Iowa Carver College of Medicine, Iowa City, IA 52242J. Stacey Klutts - Department of Pathology, University of Iowa Carver College of Medicine, 200 Hawkins Drive, Iowa City, IA 52242, United States
- Resource Type
- Journal article
- Publication Details
- Journal of microbiological methods, Vol.95(3), pp.373-378
- Publisher
- Elsevier B.V
- DOI
- 10.1016/j.mimet.2013.09.021
- PMID
- 24161898
- ISSN
- 0167-7012
- eISSN
- 1872-8359
- Language
- English
- Date published
- 12/2013
- Academic Unit
- Molecular Physiology and Biophysics; Pathology; Internal Medicine
- Record Identifier
- 9984025432302771
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