Efficient transformation of the beetle Tribolium castaneum using the Minos transposable element: quantitative and qualitative analysis of genomic integration events.
Genetic transformation in insects holds great promise as a tool for genetic manipulation in species of particular scientific, economic, or medical interest. A number of transposable elements have been tested recently as potential vectors for transformation in a range of insects. Minos is one of the most promising elements because it appears to be active in diverse species and has the capacity to carry large inserts. We report here the use of the Minos element as a transformation vector in the red flour beetle Tribolium castaneum (Coleoptera), an important species for comparative developmental and pest management studies. Transgenic G(1) beetles were recovered from 32.4% of fertile G(0)'s injected with a plasmid carrying a 3xP3-EGFP-marked transposon and in vitro synthesized mRNA encoding the Minos transposase. This transformation efficiency is 2.8-fold higher than that observed when using a plasmid helper. Molecular and genetic analyses show that several independent insertions can be recovered from a single injected parent, but that the majority of transformed individuals carry single Minos insertions. These results establish Minos as one of the most efficient vectors for genetic transformation in insects. In combination with piggyBac-based transgenesis, our work allows the introduction of sophisticated multicomponent genetic tools in Tribolium.
Pavlopoulos
Anastasios
A
Institute of Molecular Biology and Biotechnology (IMBB-FORTH), 711 10 Iraklio Crete, Greece. pavlopou@imbb.forth.gr
Berghammer
Andreas J
AJ
Averof
Michalis
M
Klingler
Martin
M
D000818:Animals; D001483:Base Sequence; D003433:Crosses, Genetic; D004251:DNA Transposable Elements / Q000235:genetics; D005260:Female; D016678:Genome; D008297:Male; D016254:Mutagenesis, Insertional; D010957:Plasmids; D014170:Transformation, Genetic; D019895:Transposases / Q000378:metabolism*; D014231:Tribolium / Q000235:genetics*
D004251:DNA Transposable Elements; C473499:Minos transposase; D019895:Transposases