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Creation of a set of 2EP lines

The goal was to set up a series of chromosomes, each carrying two EP elements that are 1-2 Mbp apart, which can be used to generate a series of recombinants between the mutation and the EP insertions for a molecular-scale mapping of the mutation relative to the SNP sites between the EP inserts. These recombinants can be identified based on eye-colour differences in animals carrying neither, one, or two of these elements. Using such stocks, it is a simple matter to isolate a set of recombinant chromosomes with an average of one recombination event every 10-20kb.

We have accomplished the following tasks:
- We selected 20 EP insertions for each of the 5 major chromosome arms. These insertions are homozygous viable, the flanking 3000bp DNA has no repetitive sequences and the adults carrying one or two EP elements can be distinguished based on their eye colours.

- All of the planned sets of 2EP stocks were generated except for chromosome arm 3R. 18 stocks for chromosome X, 18 stocks for 2L, 17 stocks for 2R, 15 stocks for 3L, and 11 stocks for 3R are now available.

- Verification of the 2EP insertions by PCR has been successfully performed with all available 2EP stocks.

With these 2EP lines, it is possible to fine-map mutations to a resolution of 10-20kb. In particular, if the location of the mutation could be narrowed down by the newly developed "tag"-array minisequencing technique to ca. 2Mbp, the 2EP lines provide a useful tool for the next, fine-mapping step. Thus, these stocks provide a further tool, which facilitates localization of genes. Consequently, one of the major rate-limiting steps in the study of Drosophila functional genomics assigning known functions to specific genes - is greatly accelerated.

The 2EP lines are freely available for all Drosophila researchers. Both the Szeged
Drosophila Transposon Stock Center and IMP/IMBA (Vienna) are keeping at least one copy of each of the verified 2EP stocks.

Informations connexes

Résultat en bref

Reported by

IMBA, Austrian Academy of Sciences
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