Mapping Drosophila mutations with molecularly defined P element insertions.
The isolation of chemically induced mutations in forward genetic screens is one of the hallmarks of Drosophila genetics. However, mapping the corresponding loci and identifying the molecular lesions associated with these mutations are often difficult and labor-intensive. Two mapping methods are most often used in flies: meiotic recombination mapping with marked chromosomes and deficiency mapping. The availability of the fly genome sequence allows the establishment and usage of molecular markers. Single-nucleotide polymorphisms have therefore recently been used to map several genes. Here we show that thousands of molecularly mapped P element insertions in fly strains that are publicly available provide a powerful alternative method to single-nucleotide polymorphism mapping. We present a strategy that allows mapping of lethal mutations, as well as viable mutations with visible phenotypes, with minimal resources. The most important unknown in using recombination rates to map at high resolution is how accurately recombination data correlate with molecular maps in small intervals. We therefore surveyed distortions of recombination rates in intervals <500 kb. We document the extent of distortions between the recombination and molecular maps and describe the required steps to map with an accuracy of <50 kb. Finally, we describe a recently developed method to determine molecular lesions in 50-kb intervals by using a heteroduplex DNA mutation detection system. Our data show that this mapping approach is inexpensive, efficient, and precise, and that it significantly broadens the application of P elements in Drosophila.
Zhai
R Grace
RG
Howard Hughes Medical Institute, Baylor College of Medicine, Houston, TX 77030, USA.
Hiesinger
P Robin
PR
Koh
Tong-Wey
TW
Verstreken
Patrik
P
Schulze
Karen L
KL
Cao
Yu
Y
Jafar-Nejad
Hamed
H
Norga
Koenraad K
KK
Pan
Hongling
H
Bayat
Vafa
V
Greenbaum
Michael P
MP
Bellen
Hugo J
HJ
D000818:Animals; D004251:DNA Transposable Elements; D004330:Drosophila / Q000235:genetics*; D008540:Meiosis / Q000235:genetics; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D011995:Recombination, Genetic
D004251:DNA Transposable Elements