Introduction of the transposable element Minos into the germ line of Drosophila melanogaster.
A transposon based on the transposable element Minos from Drosophila hydei was introduced into the genome of Drosophila melanogaster using transformation mediated by the Minos transposase. The transposon carries a wild-type version of the white gene (w) of Drosophila inserted into the second exon of Minos. Transformation was obtained by injecting the transposon into preblastoderm embryos that were expressing transposase either from a Hsp70-Minos fusion inserted into the genome via P-element-mediated transformation or from a coinjected plasmid carrying the Hsp70-Minos fusion. Between 1% and 6% of the fertile injected individuals gave transformed progeny. Four of the insertions were cloned and the DNA sequences flanking the transposon ends were determined. The "empty" sites corresponding to three of the insertions were amplified from the recipient strain by PCR, cloned, and sequenced. In all cases, the transposon has inserted into a TA dinucleotide and has created the characteristic TA target site duplication. In the absence of transposase, the insertions were stable in the soma and the germ line. However, in the presence of the Hsp70-Minos gene the Minos-w transposon excises, resulting in mosaic eyes and germ-line reversion to the white phenotype. Minos could be utilized as an alternative to existing systems for transposon tagging and enhancer trapping in Drosophila; it might also be of use as a germ-line transformation vector for non-Drosophila insects.
Loukeris
T G
TG
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, Heraklion, Greece.
Arcà
B
B
Livadaras
I
I
Dialektaki
G
G
Savakis
C
C
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D004251:DNA Transposable Elements / Q000235:genetics*; D004331:Drosophila melanogaster / Q000201:enzymology / Q000235:genetics*; D005123:Eye / Q000033:anatomy & histology; D005822:Genetic Vectors; D005854:Germ Cells; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000235:genetics; D010641:Phenotype; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic; D019895:Transposases
D004251:DNA Transposable Elements; D009713:Nucleotidyltransferases; D019895:Transposases