Rapid genetic mapping in Neurospora crassa.
Forward genetic analysis is the most broadly applicable approach to discern gene functions. However, for some organisms like the filamentous ascomycete Neurospora crassa, genetic mapping frequently represents a limiting step in forward genetic approaches. We describe an efficient method for genetic mapping in N. crassa that makes use of a modified bulked segregant analysis and PCR-based molecular markers. This method enables mapping with progeny from a single cross and requires only 90 PCR amplifications. Genetic distances between syntenic markers have been determined to ensure complete coverage of the genome and to allow interpolation of linkage data. As a result, most mutations should be mapped in less than one month to within 1-5 map units, a level of resolution sufficient to initiate map-based cloning efforts. This system also will facilitate analyses of recombination at a genome-wide level and is applicable to other perfect fungi when suitable markers are available.
Jin
Yuan
Y
Department of Biology, Texas A&M University, 3258 TAMU, College Station, TX 77843, USA.
Allan
Sabrina
S
Baber
Lauren
L
Bhattarai
Eric K
EK
Lamb
Teresa M
TM
Versaw
Wayne K
WK
D004252:DNA Mutational Analysis; D005819:Genetic Markers; D016681:Genome, Fungal / Q000235:genetics*; D009154:Mutation; D009492:Neurospora crassa / Q000235:genetics*; D020161:Physical Chromosome Mapping / Q000379:methods*; D011995:Recombination, Genetic; D012680:Sensitivity and Specificity
D005819:Genetic Markers