Transfer and expression of an organophosphate insecticide-degrading gene from Pseudomonas in Drosophila melanogaster.
The organophosphorus acid hydrolases represent a distinct class of enzymes that catalyze the hydrolysis of a variety of organophosphate substrates, including many insecticides and their structural analogues. The plasmid-borne opd gene of Pseudomonas diminuta strain MG specifies an organophosphorus acid hydrolase, a phosphotriesterase, that has been well characterized and can hydrolyze a broad spectrum of insect and mammalian neurotoxins. The in situ functioning of this enzyme in the metabolism of organophosphates has been analyzed directly in insects by transferring the opd gene into embryos of Drosophila melanogaster by P element-mediated transformation. The chromosomal locations of this stably inherited transgenic locus differed from strain to strain and demonstrated various expressivity on the whole-insect basis. Transcriptional induction of opd in one of these strains under control of the Drosophila heat shock promoter, hsp70, resulted in the synthesis of stable active enzyme that accumulated to high levels with repeated induction. The heat shock-induced synthesis of organophosphorus acid hydrolases in transgenic flies conferred enhanced resistance to toxic paralysis by the organophosphate insecticide paraoxon.
Phillips
J P
JP
Department of Molecular Biology, University of Guelph, ON, Canada.
Xin
J H
JH
Kirby
K
K
Milne
C P
CP
Krell
P
P
Wild
J R
JR
D000818:Animals; D043303:Aryldialkylphosphatase; D004331:Drosophila melanogaster / Q000187:drug effects / Q000235:genetics*; D004351:Drug Resistance / Q000235:genetics; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D006360:Heat-Shock Proteins / Q000235:genetics; D006358:Hot Temperature; D008297:Male; D010261:Paraoxon / Q000494:pharmacology*; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011549:Pseudomonas / Q000235:genetics*; D015183:Restriction Mapping; D014162:Transfection
D006360:Heat-Shock Proteins; D010744:Phosphoric Monoester Hydrolases; D043303:Aryldialkylphosphatase; D010261:Paraoxon