A set of transgenic insertion stocks derived by TE mobilization using the P-element construct P{EP}. The P{EP} construct construct carries a w+mC mini-white visible marker, Scer\UAS binding sites for the Scer\GAL4 transcriptional regulator, and bacterial sequences that allow plasmid rescue. The GAL4-UAS system allows regulated expression of genes proximate to the site of the insertion: genes properly oriented with respect to the Scer\UAS sequences can be conditionally expressed via transgene-derived Scer\GAL4 activity.
Insertion lines from this collection were mapped and assessed for inclusion in the Gene Disruption Project collection; flanking sequence data were submitted to GenBank.
Initial insertions were recovered after injection of a plasmid containing P{EP} into embryos; additional sites were recovered after exposure to the transposase-producing P{Δ2-3}99B construct on the third chromosome. Insertions were mapped to a chromosome and balanced; lines for which a single P-element was detected by Southern blot and by in situ hybridization to polytene chromosomes were characterized further.
Mobilization of the P-element occurred in animals (both males and females were used) carrying P{EP} on a second chromosome CyO balancer chromosome and P{Δ2-3}99B, a stable source of P transposase, on a marked third chromosome. 2300 independent new inserts were mapped to chromosomes X, 2 or 3 and balanced for further analysis.
Markers in stocks (as originally submitted to the BDSC):w1118.
For 2266 lines generated in the Rorth laboratory, flanking genomic sequences were obtained by sequencing of inverse PCR products, and were mapped by alignment to the genomic sequence. 374 lines were selected for inclusion in the BDGP Gene Disruption Project 2004 collection.
