Cloning and expression of an atrazine inducible cytochrome P450, CYP4G33, from Chironomus tentans (Diptera: Chironomidae) |
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Authors: | Diana K Londoño Haichuan Wang Michael J Lydy |
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Institution: | a Department of Entomology, University of Nebraska-Lincoln, 202 Plant Industry Building, Lincoln, NE 68583, USA b Universidade Federal Rural de Pernambuco, Departamento de Agronomia—Entomologia, 52171-900 Recife-PE, Brazil c USDA-ARS Grain, Forage & Bioenergy Research, Lincoln, NE 68583, USA d Department of Zoology, Fisheries and Illinois Aquaculture Center, Southern Illinois University, Carbondale, IL 62901, USA |
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Abstract: | Previous studies performed in our laboratory have measured the effect of atrazine exposure on cytochrome P450-dependent monooxygenase activity and have found increased activity in midge larvae (Chironomus tentans) as a result of atrazine exposure (1-10 ppm). Here we report the cloning and expression of a specific C. tentans CYP4 gene that is responsive to atrazine induction with an open reading frame of 1678 bp which encodes a putative protein of 559 amino acid residues. Alignments of deduced amino acid sequences with other insect P450 genes and phylogenetic analysis indicated a high degree of similarity to other insect CYP4 genes. Northern blotting analysis employing a fragment of 1200 bp from the CYP4 gene as a probe indicated that the CYP4 gene was expressed in all developmental stages, but was expressed at highest levels in late instar larvae. Additionally, over-expression of CYP4 in C. tentans exposed to atrazine (10 mg/l) confirms the ability of atrazine to induce specific P450 genes and provides insight into potential consequences of atrazine exposure in aquatic organisms. |
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Keywords: | Cytochrome P450 Atrazine Induction Chironomus tentans Expression |
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