Mapping Drosophila mutations with molecularly defined P element insertions

R. Grace Zhai, P. Robin Hiesinger, Tong Wey Koh, Patrik Verstreken, Karen L. Schulze, Yu Cao, Hamed Jafar-Nejad, Koenraad K. Norga, Hongling Pan, Vafa Bayat, Michael P. Greenbaum, Hugo J. Bellen

Research output: Contribution to journalArticle

82 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)10860-10865
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume100
Issue number19
DOIs
StatePublished - Sep 16 2003

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Genetic Recombination
Drosophila
Mutation
Diptera
Single Nucleotide Polymorphism
Nucleotide Mapping
Nucleic Acid Heteroduplexes
Chromosome Mapping
Genome
Phenotype
Genes

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Zhai, R. G., Hiesinger, P. R., Koh, T. W., Verstreken, P., Schulze, K. L., Cao, Y., ... Bellen, H. J. (2003). Mapping Drosophila mutations with molecularly defined P element insertions. Proceedings of the National Academy of Sciences of the United States of America, 100(19), 10860-10865. https://doi.org/10.1073/pnas.1832753100

Mapping Drosophila mutations with molecularly defined P element insertions. / Zhai, R. Grace; Hiesinger, P. Robin; Koh, Tong Wey; Verstreken, Patrik; Schulze, Karen L.; Cao, Yu; Jafar-Nejad, Hamed; Norga, Koenraad K.; Pan, Hongling; Bayat, Vafa; Greenbaum, Michael P.; Bellen, Hugo J.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, No. 19, 16.09.2003, p. 10860-10865.

Research output: Contribution to journalArticle

Zhai, RG, Hiesinger, PR, Koh, TW, Verstreken, P, Schulze, KL, Cao, Y, Jafar-Nejad, H, Norga, KK, Pan, H, Bayat, V, Greenbaum, MP & Bellen, HJ 2003, 'Mapping Drosophila mutations with molecularly defined P element insertions', Proceedings of the National Academy of Sciences of the United States of America, vol. 100, no. 19, pp. 10860-10865. https://doi.org/10.1073/pnas.1832753100
Zhai, R. Grace ; Hiesinger, P. Robin ; Koh, Tong Wey ; Verstreken, Patrik ; Schulze, Karen L. ; Cao, Yu ; Jafar-Nejad, Hamed ; Norga, Koenraad K. ; Pan, Hongling ; Bayat, Vafa ; Greenbaum, Michael P. ; Bellen, Hugo J. / Mapping Drosophila mutations with molecularly defined P element insertions. In: Proceedings of the National Academy of Sciences of the United States of America. 2003 ; Vol. 100, No. 19. pp. 10860-10865.
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