Systematic generation of high-resolution deletion coverage of the Drosophila melanogaster genome.
In fruit fly research, chromosomal deletions are indispensable tools for mapping mutations, characterizing alleles and identifying interacting loci. Most widely used deletions were generated by irradiation or chemical mutagenesis. These methods are labor-intensive, generate random breakpoints and result in unwanted secondary mutations that can confound phenotypic analyses. Most of the existing deletions are large, have molecularly undefined endpoints and are maintained in genetically complex stocks. Furthermore, the existence of haplolethal or haplosterile loci makes the recovery of deletions of certain regions exceedingly difficult by traditional methods, resulting in gaps in coverage. Here we describe two methods that address these problems by providing for the systematic isolation of targeted deletions in the D. melanogaster genome. The first strategy used a P element-based technique to generate deletions that closely flank haploinsufficient genes and minimize undeleted regions. This deletion set has increased overall genomic coverage by 5-7%. The second strategy used FLP recombinase and the large array of FRT-bearing insertions described in the accompanying paper to generate 519 isogenic deletions with molecularly defined endpoints. This second deletion collection provides 56% genome coverage so far. The latter methodology enables the generation of small custom deletions with predictable endpoints throughout the genome and should make their isolation a simple and routine task.
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Annette L
AL
Parks
Kevin R
KR
Cook
Marcia
M
Belvin
Nicholas A
NA
Dompe
Robert
R
Fawcett
Kari
K
Huppert
Lory R
LR
Tan
Christopher G
CG
Winter
Kevin P
KP
Bogart
Jennifer E
JE
Deal
Megan E
ME
Deal-Herr
Deanna
D
Grant
Marie
M
Marcinko
Wesley Y
WY
Miyazaki
Stephanie
S
Robertson
Kenneth J
KJ
Shaw
Mariano
M
Tabios
Valentina
V
Vysotskaia
Lora
L
Zhao
Rachel S
RS
Andrade
Kyle A
KA
Edgar
Elizabeth
E
Howie
Keith
K
Killpack
Brett
B
Milash
Amanda
A
Norton
Doua
D
Thao
Kellie
K
Whittaker
Millicent A
MA
Winner
Lori
L
Friedman
Jonathan
J
Margolis
Matthew A
MA
Singer
Casey
C
Kopczynski
Daniel
D
Curtis
Thomas C
TC
Kaufman
Gregory D
GD
Plowman
Geoffrey
G
Duyk
Helen L
HL
Francis-Lang
D000818:Animals; D004251:DNA Transposable Elements; D004331:Drosophila melanogaster / Q000235:genetics*; D016678:Genome; D016254:Mutagenesis, Insertional; D017384:Sequence Deletion
D004251:DNA Transposable Elements