Mobility assays confirm the broad host-range activity of the Minos transposable element and validate new transformation tools.
Autor(es): Klinakis A G; Loukeris T G; Pavlopoulos A; Savakis C
Resumo: Fast and reliable methods for assessing the mobility of the transposable element Minos have been developed. These methods are based on the detection of excision and insertion of Minos transposons from and into plasmids which are co-introduced into cells. Excision is detected by polymerase chain reaction (PCR) with appropriate primers. Transposition is assayed by marker rescue in Escherichia coli, using a transposon plasmid that carries a tetracycline resistance gene and a target plasmid carrying a gene that can be selected against in E. coli. Using both assays, Minos was shown to transpose in Drosophila melanogaster cells and embryos, and in cultured cells of a mosquito, Aedes aegypti, and a lepidopteran, Spodoptera frugiperda. In all cases, mobility was dependent on the presence of exogenously supplied transposase, and both excision and transposition were precise. The results indicate that Minos can transpose in heterologous insect species with comparable efficiencies and therefore has the potential to be used as a transgenesis vector for diverse species.
Palavras-Chave: Transposable element; Minos insect transgenesis; Transformation vector; Transposition assay
Imprenta: Insect Molecular Biology, v. 9, n. 3, p. 269-275, 2000
Identificador do objeto digital: 10.1046/j.1365-2583.2000.00183.x
Descritores: Aedes aegypti - Cell ; Aedes aegypti - DNA ; Aedes aegypti - Genome ; Aedes aegypti - Molecular Structure ; Aedes aegypti - Pathogenesis ; Aedes aegypti - Molecular methods ; Aedes aegypti - PCR detection
Data de publicação: 2000