Microbiology and Kluyver Centre for Genomics of Industrial Fermentations, Institute of Biomembranes, Utrecht University, Padualaan 8, Utrecht, the Netherlands.
FEMS Microbiol Lett. 2010 Sep 1;310(1):91-5. doi: 10.1111/j.1574-6968.2010.02052.x. Epub 2010 Jul 2.
Schizophyllum commune is the only mushroom-forming fungus in which targeted gene deletions by homologous recombination have been reported. However, these deletions occur with a low frequency. To overcome this, the ku80 gene of S. commune was deleted. This gene is involved in the nonhomologous end-joining system for DNA repair. The Deltaku80 strain was not affected in growth and development. However, the transformation efficiency was reduced up to 100-fold. This was accompanied by a strong increase in the relative number of transformants with a homologous integration of a knockout construct. Genes sc15, jmj3 and pri2 were deleted in the Deltaku80 strain. In total, seven out of 10 transformants showed a gene deletion. This frequency will facilitate a systematic analysis of gene function in S. commune.
栓菌是唯一一种被报道可以通过同源重组进行靶向基因缺失的蕈类真菌。然而，这些缺失的发生频率很低。为了解决这个问题，我们删除了栓菌的 ku80 基因。该基因参与 DNA 修复的非同源末端连接系统。Deltaku80 菌株在生长和发育上不受影响。然而，转化效率降低了多达 100 倍。与此同时，同源整合敲除构建体的转化子的相对数量大大增加。sc15、jmj3 和 pri2 基因在 Deltaku80 菌株中被删除。在总共 10 个转化子中，有 7 个显示出基因缺失。这个频率将有助于系统地分析栓菌中的基因功能。
