The availability of an organism’s genome sequence is useful for improving downstream applications such as large-scale biofuel production, but it is only the first step on this path. In the case of the fungus Trichoderma reesei, whose genome sequence was published by the DOE JGI in 2008, the cellulases in T. reesei have multiple industrial applications, but fungal strains have traditionally been developed by random mutagenesis.
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| T. reesei (Image by Irma Salovuori, VTT Biotechnology) |
A functional analysis of the predicted genes identified in the genome would be useful for a more directed approach to strain development, but the time and effort needed to create a gene knockout library, which can require thousands of experiments is an obstacle that needs to be overcome.
To speed up the process and more efficiently use these genomic resources, a team of researchers including DOE JGI’s Scott Baker at Pacific Northwest National Laboratory have developed a construction kit that can be applied in a high throughput approach to knock out genes in T. reesei in order to analyze gene functions. Their process, which involves the method of yeast mediated recombination for deletion vector construction, is described in the January 2, 2012 issue of Biotechnology for Biofuels.
“Our streamlined system for knockout-construction along with the convenient and easy to use method to create multiple mutants by crossing can serve as a starting point for large scale functional analysis studies aimed to uncover the hidden treasures of the biotechnological workhorse T. reesei,” Baker and his colleagues wrote.