Insertion lines from this collection were assessed for inclusion in the BDGP Gene Disruption Project collection.
A set of mutant stocks derived by insertional mutagenesis using the P-element construct P{hsneo}; most lines have a lethal or sterile phenotype. The P{hsneo} construct carries a neomycin-resistance gene with a heat-shock promoter (allowing chemical selection in flies using G418), and bacterial sequences that allow plasmid rescue. First dataset of this type.
A genetic screen allowing selection of new autosomal insertions was used. Parental males carried a P{hsneo} insertion on the X chromosome and P{JS}, a transposase-producing "jumpstarter" construct, on the third chromosome. They were crossed to females carrying a marked third chromosome and their progeny were exposed to G418. An F1 male with a new autosomal insertion was able to survive G418 selection, allowing isolation of these relatively rare transposition events. Stable stocks were established, using a single F1 male from a given parental male to avoid premeiotic clusters.
General location of each insertion was determined by in situ hybridization to polytene chromosomes. Using an array of characterized autosomal deficiencies, in many cases it was possible to test for complementation with an appropriate deficiency; failure to complement resulted in the line being characterized as "verified." Flanking genomic sequence was isolated by plasmid rescue (primarily) or by inverse PCR. Original collection consisted of 24 validated lines.
Nine lines were selected for inclusion in the BDGP Gene Disruption Project 2004 collection.
