Targeted gene deletion in Aspergillus fumigatus using microbial machinery and a recyclable marker

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• 作者：Joshua B. Kieler ; Khanh L. Duong ; W. Scott Moye-Rowley ; J. Stacey Klutts
• 关键词：Aspergillus fumigatus ; Gene deletion ; Agrobacterium ; Amylase
• 刊名：Journal of Microbiological Methods
• 出版年：December, 2013
• 年：2013
• 卷：95
• 期：3
• 页码：373-378
• 全文大小：724 K

文摘

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 (12 weeks to 3 weeks) using minimal human labor, and we demonstrate that it can be used to efficiently generate multiple gene deletions within a single strain.