Journal of Economic Entomology (2007) 100, 946-953

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Dolnapa Kaewpa, Soamrutai Boonsuepsakul and Pornpimol Rongnoparut (2007)
Functional expression of mosquito NADPH-cytochrome P450 reductase in Escherichia coli
Journal of Economic Entomology 100 (3), 946-953
Abstract: A complete NADPH-cytochrome P450 reductase (CPR) cDNA was isolated from Anopheles minimus Theobald mosquitoes by using reverse transcription-polymerase chain reaction-based methods. The complete cDNA contains 2,040 bp encoding the protein of 679 amino acids, with a calculated molecular mass of 77.3 kDa. The deduced amino acid sequence had a typical feature of CPR by possessing conserved domains involved in binding of flavin mononucleotide, flavin adenine dinucleotide, and NADPH cofactors. The complete CPR cDNA was expressed as 6x His-tagged fusion protein in both membrane and cytosolic fractions in Escherichia coli, both fractions contained NADPH-cytochrome c reducing activity. The membrane-bound form containing N-terminal membrane anchor was subjected to purification, and K m values were determined for NADPH and cytochrome c. The purified CPR enzyme was functionally active, as demonstrated by its ability to support CYP6AA3-mediated metabolism in the reconstituted reaction in vitro. Initial test suggested that CYP6AA3 could play a role in deltamethrin metabolism.
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Research topic(s) for pests/diseases/weeds:
pesticide resistance of pest
pesticide resistance of pest

Pest and/or beneficial records:

Beneficial Pest/Disease/Weed Crop/Product Country Quarant.

Anopheles minimus