title: Analysis of novel alleles of brother of tout-velu, the drosophila ortholog of human EXTL3 using a newly developed FRT42D ovoDchromosome.
abstract: The FLP/FRT system permits rapid phenotypic screening of homozygous lethal mutations in the context of a viable mosaic fly. Combining this system with ovoDdominant female-sterile transgenes enables efficient production of embryos derived from mutant germline clones lacking maternal contribution from a gene of interest. Two distinct sets of FRT chromosomes, carrying either the mini-white (w+ mW.hs), or rosy (ry+) and neomycin (neoR) transgenes are in common use. Parallel ovoDlines were developed using w+ mW.hsFRT insertions on the X and chromosomes 2R and 3L, as well as ry+, neoRFRT insertions on 2L and 3R. Consequently, mutations isolated on the X, 2R and 3L chromosomes in a ry+, neoRFRT background, are not amenable to germline clonal analysis without labor-intensive recombination onto chromosome arms containing a w+ mW.hsFRT. Here we report the creation of a new ovoDline for the ry+, neoRFRT insertion at position FRT42D on chromosome 2R, through induced recombination in males. To establish the developmental relevance of this reagent we characterized the maternal-effect phenotypes of novel brother of tout-velu alleles generated on an FRT42D chromosome in a somatic mosaic screen. We find that an apparent null mutation that causes severe defects in somatic tissues has a much milder effect on embryonic patterning, emphasizing the necessity of analyzing mutant phenotypes at multiple developmental stages.
authors.affiliation: Department of Developmental and Cell Biology, University of California Irvine, Irvine, CA, 92697.
authors.forename: Ernesto
authors.initials: E
authors.lastname: Lujan
authors.affiliation: Department of Developmental and Cell Biology, University of California Irvine, Irvine, CA, 92697.|Developmental Biology Center, University of California Irvine, Irvine, CA, 92697.
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: Bornemann
authors.affiliation: Department of Biology, ETH Zurich, Institute of Molecular Systems Biology, Auguste-Piccard-Hof 1, Zurich, 8093, Switzerland.
authors.forename: Carmen
authors.initials: C
authors.lastname: Rottig
authors.affiliation: Department of Developmental and Cell Biology, University of California Irvine, Irvine, CA, 92697.
authors.forename: Brian A
authors.initials: BA
authors.lastname: Bayless
authors.affiliation: Department of Biology, ETH Zurich, Institute of Molecular Systems Biology, Auguste-Piccard-Hof 1, Zurich, 8093, Switzerland.
authors.forename: Hugo
authors.initials: H
authors.lastname: Stocker
authors.affiliation: Department of Biology, ETH Zurich, Institute of Molecular Systems Biology, Auguste-Piccard-Hof 1, Zurich, 8093, Switzerland.
authors.forename: Ernst
authors.initials: E
authors.lastname: Hafen
authors.affiliation: Department of Developmental and Cell Biology, University of California Irvine, Irvine, CA, 92697.|Developmental Biology Center, University of California Irvine, Irvine, CA, 92697.
authors.forename: Kavita
authors.initials: K
authors.lastname: Arora
authors.affiliation: Department of Developmental and Cell Biology, University of California Irvine, Irvine, CA, 92697.|Developmental Biology Center, University of California Irvine, Irvine, CA, 92697.
authors.forename: Rahul
authors.initials: R
authors.lastname: Warrior
mesh_terms: D000483:Alleles; D000818:Animals; D019521:Body Patterning / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics* / Q000254:growth & development; D005260:Female; D018507:Gene Expression Regulation, Developmental; D018095:Germ-Line Mutation / Q000235:genetics; D006801:Humans; D007247:Infertility, Female / Q000235:genetics*; D008297:Male; D009030:Mosaicism / Q000196:embryology; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics*; D010641:Phenotype; D000072020:Synthetic Lethal Mutations / Q000235:genetics; D019076:Transgenes
chemical_list: D029721:Drosophila Proteins; C110774:EXTL3 protein, human; C487132:Botv protein, Drosophila; D017351:N-Acetylglucosaminyltransferases
keywords: Drosophila FRT42D ovoD; EXTL3; HSPG; N-acetylglucosamine transferase-I/II; botv; brother of tout-velu; germline clonal analysis; heparan sulfate proteoglycan