Retrotransposition of a marked Drosophila line-like I element in cells in culture.

Silke Jensen (1) , L. Cavarec (1) , O. Dhellin (1) , T. Heidmann (1)

1 REERES (UMR 8122) - Rétrovirus endogènes et éléments rétroides des eucaryotes supérieurs

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.

Domaines: Génomique, Transcriptomique et Protéomique [q-bio.GN]

Silke Jensen, L. Cavarec, O. Dhellin, T. Heidmann. Retrotransposition of a marked Drosophila line-like I element in cells in culture.. Nucleic Acids Research, 1994, 22 (8), pp.1484-1488. ⟨10.1093/nar/22.8.1484⟩. ⟨hal-01925042⟩