Genetics · 1999 · 44 citations · 40 references
Life SciencesGeneticsMolecular GeneticsGene ExpressionGenomicsInsect GeneticsHorizontal Gene TransferOogenesisHybrid DysgenesisImmunofluorescence ImagingIndirect Immunofluorescence AssayLong Terminal Repeat RetrotransposonsNon-long Terminal Repeat RetrotransposonsRetrotranspositionTransposition EventsTransposition Mechanisms
To study the expression of the I factor, a non-long-terminal-repeat retrotransposon responsible for I-R hybrid dysgenesis in Drosophila melanogaster, we have tagged the ORF1 protein (ORF1p) by inserting the HA epitope in its N-terminal region. In transgenic flies, this modification is compatible with a high rate of autonomous transposition and allows direct estimation of the transposition frequency. I factor transposes in the germline of females (SF) that are daughters from crosses between I strain males (which contain active copies of the I factor) and R strain females (which do not). We analyzed the expression pattern of ORF1p by indirect immunofluorescence. Its expression correlates with retrotransposition. During oogenesis ORF1p appears unexpectedly as a cytoplasmic product, which accumulates with a specific pattern into the oocyte. A comparison of the expression patterns under conditions that modify the transposing activity of the element clarifies some aspects of I-factor functioning in the transposition process.
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