Transformation of Stomoxys calcitrans with a Hermes gene vector.
The ability of the Hermes transposable element to function as a germ line transformation vector was tested in the stable fly, Stomoxys calcitrans. Plasmid-based transposable element mobility assays indicated moderate mobility of Hermes in this species. Germline transformants were created using a Hermes element containing the enhanced green fluorescent protein (EGFP) under the regulatory control of the promoter from Actin5C gene of Drosophila melanogaster. Approximately 4% of the fifty-five adults that developed from the 1903 G(0) embryos injected with the vector produced transgenic progeny. In the four transgenic lines established, the EGFP expression pattern was distinctly nonuniform and levels of expression were low. Promoters other than the one from the Actin5C gene of D. melanogaster should be considered for widespread, constitutive expression. All transgenic lines contained multiple (2-4) integrated Hermes elements. Hermes integration events occurred through a canonical cut-and-paste mechanism.
O'Brochta
D A
DA
Center for Agricultural Biotechnology, University of Maryland Biotechnology Institute, College Park, MD 20742-4450, USA. obrochta@umbi.umd.edu
Atkinson
P W
PW
Lehane
M J
MJ
D000818:Animals; D030801:Animals, Genetically Modified; D015139:Blotting, Southern; D004251:DNA Transposable Elements; D005260:Female; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D009117:Muscidae / Q000196:embryology / Q000235:genetics*; D010063:Ovum; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014170:Transformation, Genetic
D004251:DNA Transposable Elements; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins