title: Retrotransposition of a marked Drosophila line-like I element in cells in culture.
abstract: We have marked a Drosophila transposable element--the LINE-like I element--with an intron-containing indicator gene inserted in place of a large deletion in the I element second ORF encompassing the reverse transcriptase domain, and this marked element was placed downstream to a potent actin promoter. An expression vector for the I element ORFs was also constructed, under the same heterologous promoter. The indicator gene contains a lacZ reporter gene the expression of which is conditioned by retrotransposition of the marked element, thus allowing detection of transposition events by testing for either beta-galactosidase expression or occurrence of spliced DNA molecules. The marked I element was introduced into Drosophila melanogaster cells in culture by transfection. Spliced DNA copies of the marked element and specifically stained beta-galactosidase-expressing cells were detected only upon co-transfection with the I expression vector, thus indicating that an ORF2-deleted element can be complemented in trans for transposition. This simple assay for retrotransposition in Drosophila cells in culture provides a tool for the rapid analysis of the mechanism of I transposition in its cis and trans sequence requirements.
authors.affiliation: Unités de Physicochimie et Pharmacologie des Macromolécules Biologiques, CNRS, Villejuif, France.
authors.forename: S
authors.initials: S
authors.lastname: Jensen
authors.forename: L
authors.initials: L
authors.lastname: Cavarec
authors.forename: O
authors.initials: O
authors.lastname: Dhellin
authors.forename: T
authors.initials: T
authors.lastname: Heidmann
mesh_terms: D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004247:DNA; D004251:DNA Transposable Elements; D004331:Drosophila melanogaster / Q000166:cytology / Q000235:genetics*; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D014162:Transfection
chemical_list: D004251:DNA Transposable Elements; D004247:DNA