Cloning and expression of an atrazine inducible cytochrome P450, CYP4G33, from Chironomus tentans (Diptera: Chironomidae)

Authors

Diana K. Londono, University of Nebraska-Lincoln
Herbert A.A. Siqueira, Universidade Federal Rural de Pernambuco, Departamento de Agronomia—Entomologia, 52171-900 Recife-PE, Brazil
Haichuan Wang, University of Nebraska-LincolnFollow
Gautam Sarath, University of Nebraska-LincolnFollow
Michael J. Lydy, Southern Illinois UniversityFollow
Blair D. Siegfried, University of Nebraska-LincolnFollow

ORCID IDs

Blair D. Siegfried

Document Type

Article

Date of this Version

8-27-2007

Comments

Published in Pesticide Biochemistry and Physiology 89 (2007) 104–110.

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.