Characterisation of Drosophila Ubx CPTI000601 and hth CPTI000378 protein trap lines

ScientificWorldJournal. 2014:2014:191535. doi: 10.1155/2014/191535. Epub 2014 Oct 15.

Abstract

In Drosophila, protein trap strategies provide powerful approaches for the generation of tagged proteins expressed under endogenous control. Here, we describe expression and functional analysis to evaluate new Ubx and hth protein trap lines generated by the Cambridge Protein Trap project. Both protein traps exhibit spatial and temporal expression patterns consistent with the reported endogenous pattern in the embryo. In imaginal discs, Ubx-YFP is expressed throughout the haltere and 3rd leg imaginal discs, while Hth-YFP is expressed in the proximal regions of haltere and wing discs but not in the pouch region. The Ubx (CPTI000601) line is semilethal as a homozygote. No T3/A1 to T2 transformations were observed in the embryonic cuticle or the developing midgut. The homozygous survivors, however, exhibit a weak haltere phenotype with a few wing-like marginal bristles on the haltere capitellum. Although hth (CPTI000378) is completely lethal as a homozygote, the hth (CPTI000378) /hth (C1) genotype is viable. Using a hth deletion (Df(3R)BSC479) we show that hth (CPTI000378) /Df(3R)BSC479 adults are phenotypically normal. No transformations were observed in hth (CPTI000378), hth (CPTI000378) /hth (C1), or hth (CPTI000378) /Df(3R)BSC479 embryonic cuticles. We have successfully characterised the Ubx-YFP and Hth-YFP protein trap lines demonstrating that the tagged proteins show appropriate expression patterns and produce at least partially functional proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Drosophila Proteins / deficiency
  • Drosophila Proteins / genetics*
  • Drosophila Proteins / metabolism
  • Drosophila melanogaster / genetics*
  • Drosophila melanogaster / growth & development
  • Drosophila melanogaster / metabolism
  • Embryo, Nonmammalian
  • Founder Effect
  • Gene Deletion
  • Gene Expression Regulation, Developmental*
  • Genes, Reporter
  • Genetic Engineering
  • Genotype
  • Homeodomain Proteins / genetics*
  • Homeodomain Proteins / metabolism
  • Homozygote
  • Imaginal Discs / growth & development
  • Imaginal Discs / metabolism
  • Intestinal Mucosa / metabolism
  • Intestines / growth & development
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / metabolism
  • Phenotype
  • Recombinant Fusion Proteins / genetics*
  • Recombinant Fusion Proteins / metabolism
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism

Substances

  • Bacterial Proteins
  • Drosophila Proteins
  • Homeodomain Proteins
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • Transcription Factors
  • Ubx protein, Drosophila
  • hth protein, Drosophila
  • yellow fluorescent protein, Bacteria