Analysis of novel alleles of brother of tout-velu, the drosophila ortholog of human EXTL3 using a newly developed FRT42D ovoD chromosome

Genesis. 2016 Nov;54(11):573-581. doi: 10.1002/dvg.22981. Epub 2016 Oct 3.

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 ovoD dominant 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 ovoD lines were developed using w + mW.hs FRT insertions on the X and chromosomes 2R and 3L, as well as ry+ , neoR FRT insertions on 2L and 3R. Consequently, mutations isolated on the X, 2R and 3L chromosomes in a ry+ , neoR FRT background, are not amenable to germline clonal analysis without labor-intensive recombination onto chromosome arms containing a w + mW.hs FRT. Here we report the creation of a new ovoD line for the ry+ , neoR FRT 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.

Keywords: Drosophila FRT42D ovoD; EXTL3; HSPG; N-acetylglucosamine transferase-I/II; botv; brother of tout-velu; germline clonal analysis; heparan sulfate proteoglycan.

MeSH terms

  • Alleles
  • Animals
  • Body Patterning / genetics*
  • Drosophila Proteins / genetics*
  • Drosophila melanogaster / genetics*
  • Drosophila melanogaster / growth & development
  • Female
  • Gene Expression Regulation, Developmental
  • Germ-Line Mutation / genetics
  • Humans
  • Infertility, Female / genetics*
  • Male
  • Mosaicism / embryology
  • N-Acetylglucosaminyltransferases / genetics*
  • Phenotype
  • Synthetic Lethal Mutations / genetics
  • Transgenes

Substances

  • Drosophila Proteins
  • EXTL3 protein, human
  • Botv protein, Drosophila
  • N-Acetylglucosaminyltransferases