Florea Simona, Machado Caroline, Andreeva Kalina, Schardl Christopher L
Department of Plant Pathology, UK-Advanced Genetic Technologies Center, University of Kentucky, Lexington, KY, USA.
Methods Mol Biol. 2012;824:371-9. doi: 10.1007/978-1-61779-433-9_19.
Interspecific hybridization is a common evolutionary process for the many epichloid fungi that consequently possess multiple gene copies accumulated from their parental strains. Serial gene manipulations in such strains are impeded by the limited availability of selectable resistance marker genes. Therefore, we developed a method for marker elimination suitable for a range of filamentous fungi that allows the reuse of the same marker for successive manipulations, and can also generate gene knockout mutants free of any foreign genes. For epichloae, the complete elimination of the marker gene from the genome would mitigate public concerns and regulatory hurdles to the use of such fungal strains in field experiments.
种间杂交是许多表观真菌常见的进化过程，因此这些真菌拥有从其亲本菌株积累的多个基因拷贝。此类菌株中的系列基因操作受到可选择抗性标记基因可用性有限的阻碍。因此，我们开发了一种适用于多种丝状真菌的标记消除方法，该方法允许同一标记重复用于后续操作，还能产生不含任何外源基因的基因敲除突变体。对于内生真菌而言，从基因组中完全消除标记基因将减轻公众对在田间试验中使用此类真菌菌株的担忧和监管障碍。
