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PMID: 10049925 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Conversion of lacZ enhancer trap lines to GAL4 lines using targeted transposition in Drosophila melanogaster.

Genetics ·Vol. 151 ·No. 3 ·1999-03-00 ·Pages 1093-101

Sepp KJ, Auld VJ

Abstract

Since the development of the enhancer trap technique, many large libraries of nuclear localized lacZ P-element stocks have been generated. These lines can lend themselves to the molecular and biological characterization of new genes. However they are not as useful for the study of development of cellular morphologies. With the advent of the GAL4 expression system, enhancer traps have a far greater potential for utility in biological studies. Yet generation of GAL4 lines by standard random mobilization has been reported to have a low efficiency. To avoid this problem we have employed targeted transposition to generate glial-specific GAL4 lines for the study of glial cellular development. Targeted transposition is the precise exchange of one P element for another. We report the successful and complete replacement of two glial enhancer trap P[lacZ, ry+] elements with the P[GAL4, w+] element. The frequencies of transposition to the target loci were 1.3% and 0.4%. We have thus found it more efficient to generate GAL4 lines from preexisting P-element lines than to obtain tissue-specific expression of GAL4 by random P-element mobilization. It is likely that similar screens can be performed to convert many other P-element lines to the GAL4 system.

MeSH Terms
Animals Blotting, Southern DNA Transposable Elements Drosophila melanogaster/genetics Embryo, Nonmammalian/immunology,ultrastructure Enhancer Elements, Genetic Gene Library Genetic Testing In Situ Hybridization Lac Operon/genetics Models, Biological Models, Genetic Molecular Biology/methods Neuroglia/immunology Transcription Factors/genetics
Chemicals
DNA Transposable Elements Transcription Factors
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Sepp K J
Department of Zoology, University of British Columbia, Vancouver, British Columbia, V6T 1Z4, Canada.
Auld V J
References (24)
24 references, click to expand
  1. Molecular cloning of sequences from a Drosophila RNA polymerase II locus by P element transposon tagging.
    Cell. 1982 Dec;31(3 Pt 2):585-92 PMID: 6297774
  2. Homology requirements for targeting heterologous sequences during P-induced gap repair in Drosophila melanogaster.
    Genetics. 1997 Oct;147(2):689-99 PMID: 9335605
  3. Genetic instability in Drosophila melanogaster: P-element mutagenesis by gene conversion.
    Proc Natl Acad Sci U S A. 1988 Sep;85(17):6455-9 PMID: 2842772
  4. Embryonic development of axon pathways in the Drosophila CNS. I. A glial scaffold appears before the first growth cones.
    J Neurosci. 1989 Jul;9(7):2402-11 PMID: 2746335
  5. Searching for pattern and mutation in the Drosophila genome with a P-lacZ vector.
    Genes Dev. 1989 Sep;3(9):1273-87 PMID: 2558049
  6. P-element-mediated enhancer detection: a versatile method to study development in Drosophila.
    Genes Dev. 1989 Sep;3(9):1288-300 PMID: 2558050
  7. P-element-mediated enhancer detection: an efficient method for isolating and characterizing developmentally regulated genes in Drosophila.
    Genes Dev. 1989 Sep;3(9):1301-13 PMID: 2558051
  8. High-frequency P element loss in Drosophila is homolog dependent.
    Cell. 1990 Aug 10;62(3):515-25 PMID: 2165865
  9. The midline of the Drosophila central nervous system: a model for the genetic analysis of cell fate, cell migration, and growth cone guidance.
    Cell. 1991 Feb 22;64(4):801-15 PMID: 1997208
  10. The diversity and pattern of glia during axon pathway formation in the Drosophila embryo.
    Glia. 1991;4(2):205-13 PMID: 1827779
  11. Targeted gene replacement in Drosophila via P element-induced gap repair.
    Science. 1991 Sep 6;253(5024):1110-7 PMID: 1653452
  12. Extra sequences found at P element excision sites in Drosophila melanogaster.
    Mol Gen Genet. 1992 Mar;232(1):17-23 PMID: 1313147
  13. Genes that control neuromuscular specificity in Drosophila.
    Cell. 1993 Jun 18;73(6):1137-53 PMID: 8513498
  14. P-element-induced interallelic gene conversion of insertions and deletions in Drosophila melanogaster.
    Mol Cell Biol. 1993 Nov;13(11):7006-18 PMID: 8413290
  15. Efficient copying of nonhomologous sequences from ectopic sites via P-element-induced gap repair.
    Mol Cell Biol. 1994 Mar;14(3):1613-25 PMID: 8114699
  16. The Ets transcription factors encoded by the Drosophila gene pointed direct glial cell differentiation in the embryonic CNS.
    Cell. 1994 Jul 15;78(1):149-60 PMID: 8033206
  17. Targeting of an enhancer trap to vestigial.
    Dev Biol. 1994 Sep;165(1):290-3 PMID: 8088446
  18. Enhancer-trap targeting at the Broad-Complex locus of Drosophila melanogaster.
    Genes Dev. 1995 May 1;9(9):1137-48 PMID: 7744254
  19. Gliotactin, a novel transmembrane protein on peripheral glia, is required to form the blood-nerve barrier in Drosophila.
    Cell. 1995 Jun 2;81(5):757-67 PMID: 7539719
  20. glial cells missing: a genetic switch that controls glial versus neuronal fate.
    Cell. 1995 Sep 22;82(6):1013-23 PMID: 7553843
  21. Targeted ablation of glia disrupts axon tract formation in the Drosophila CNS.
    Development. 1995 Nov;121(11):3703-12 PMID: 8582282
  22. Distinct expression patterns detected within individual tissues by the GAL4 enhancer trap technique.
    Genome. 1996 Feb;39(1):174-82 PMID: 8851804
  23. Efficient gap repair in Drosophila melanogaster requires a maximum of 31 nucleotides of homologous sequence at the searching ends.
    Mol Cell Biol. 1997 Feb;17(2):627-34 PMID: 9001216
  24. Characterization and cloning of fasciclin III: a glycoprotein expressed on a subset of neurons and axon pathways in Drosophila.
    Cell. 1987 Mar 27;48(6):975-88 PMID: 3548998
Article Info
Journal
Genetics
Abbr.
Genetics
ISSN
0016-6731
Published
1999-03-00
Pages
1093-101
Language
English
Region
United States
NLM ID
0374636
PMCID
PMC1460539
Subset
IM
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