A set of mutant stocks derived by insertional mutagenesis using the P-element construct P{lacW}; most lines have a lethal or sterile phenotype. The P{lacW} construct carries a w+mC mini-white visible marker, Ecol\lacZ enhancer trap sequences, and bacterial sequences that allow plasmid rescue (FBrf0049800).
Insertion lines from this collection were assessed for inclusion in the Gene Disruption Project collection.
A genetic screen allowing selection of autosomal insertions was used. Parental males carried a P{lacW} insertion on the X chromosome and the transposase-producing P{Δ2-3}99B construct on a marked third chromosome; they were crossed to w[-] females. F1 males that exhibited a mini-white phenotype were presumed to carry a new autosomal insertion; these were mated individually to determine the chromosomal location of the transposed insertion. Unless they differed significantly in eye color, multiple F1 males from the same parental vial were considered to share a common insertion (via a pre-mieotic cluster). Each line was tested for lethality and stable stocks established.
A genetic screen for selection of second chromosome insertions was used. Parental males carried a P{lacW} insertion on the X chromosome and the transposase-producing P{Δ2-3}99B construct on a marked third chromosome; they were crossed en masse to w[-] females. F1 males that exhibited a mini-white phenotype were presumed to carry a new autosomal insertion; these were mated individually to determine the chromosomal location of the transposed insertion. Stable stocks were established for lethal and semi-lethal insertions on II. Pre-meiotic clusters were identified by complementation analyses between insertions arising in the same original cross.
A genetic screen allowing selection of autosomal insertions was used, as described in FBrf0049800.
An aggregate collection of lines produced in multiple different laboratories (see below). Additional lines contributed by M. Scott and M. Fuller (unpublished); no further information provided. From the screen described inFBrf0064397, only third chromosome insertions were used for this dataset.
General location of each insertion was determined by in situ hybridization to polytene chromosomes. Insertions that mapped to the same cytological location were tested for complementation. 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." For each line flanking genomic sequence was isolated by plasmid rescue (primarily) or by inverse PCR. Original collection consisted of 493 validated lines.
378 lines were selected for inclusion in the BDGP Gene Disruption Project 2004 collection.
