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Relative Efficiency Of Split-Marker Versus Double-Crossover Replacement Protocols For Production Of Deletion Mutants In Strain Ph-1 Of Fusarium Graminearum, Sladana Bec, Gabdiel E. Yulfo-Soto, Lisa J. Vaillancourt
Relative Efficiency Of Split-Marker Versus Double-Crossover Replacement Protocols For Production Of Deletion Mutants In Strain Ph-1 Of Fusarium Graminearum, Sladana Bec, Gabdiel E. Yulfo-Soto, Lisa J. Vaillancourt
Fungal Genetics Reports
The split-marker (SM) protocol has become a popular method for production of knockout mutations in fungi. We used Southern hybridization to compare the performance and efficiency of the SM protocol with the more traditional double-crossover intact marker (IM) method for creating deletions of the mating type genes in Fusarium graminearum. Both methods successfully produced knockouts at a rate of between 24 and 75%: the SM method produced mutants more efficiently for larger constructs (>1 kb), but it was similar to IM for a smaller construct that was 865 bp. Both methods also produced strains with additional ectopic integrations …