Identification, characterization, and expression of a novel P450 gene encoding CYP6AE25 from the Asian corn borer, Ostrinia furnacalis
1Key Laboratory for Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience and
Biotechnology, Chinese Academy of Tropical Agricultural Science, Haikou, Hainan 571101, P. R. China
2Key Laboratory for Biological Control of Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100081, P.R. China
3Key Laboratory of Heilongjiang Microbiology, College of Life Sciences, Heilongjiang University, Harbin, Heilongjiang
150080, P. R. China
4College of Bio-safety Science and Technology, Hunan Agricultural University, Changsha, Hunan410128, P. R. China
Abstract
An allele of the cytochrome P450 gene, CYP6AE14, named CYP6AE25 (GenBank accession no. EU807990) was isolated from the Asian corn borer, Ostrinia furnacalis (Guenée) (Lepidoptera: Pyralidae) by RT-PCR. The cDNA sequence of CYP6AE25 is 2315 bp in length and contains a 1569 nucleotides open reading frame encoding a putative protein with 523 amino acid residues and a predicted molecular weight of 59.95 kDa and a theoretical pI of 8.31. The putative protein contains the classic heme-binding sequence motif F××G×××C×G (residues 451-460) conserved among all P450 enzymes as well as other characteristic motifs of all cytochrome P450s. It shares 52% identity with the previously published sequence of CYP6AE14 (GenBank accession no. DQ986461) from Helicoverpa armigera. Phylogenetic analysis of amino acid sequences from members of various P450 families indicated that CYP6AE25 has a closer phylogenetic relationship with CYP6AE14 and CYP6B1 that are related to metabolism of plant allelochemicals, CYP6D1 which is related to pyrethroid resistance and has a more distant relationship to CYP302A1 and CYP307A1 which are related to synthesis of the insect molting hormones. The expression level of the gene in the adults and immature stages of O. furnacalis by quantitative real-time PCR revealed that CYP6AE25 was expressed in all life stages investigated. The mRNA expression level in 3rd instar larvae was 12.8- and 2.97-fold higher than those in pupae and adults, respectively. The tissue specific expression level of CYP6AE25 was in the order of midgut, malpighian tube and fatty body from high to low but was absent in ovary and brain. The analysis of the CYP6AE25 gene using bioinformatic software is discussed.
Keywords: cytochrome P450, real-time PCR, bioinformatics
Correspondence:
ayuliang1017@126.com,
bkulyemahesh@gmail.com,
cyangfshan@126.com,
dxiaoluoluo@gmail.com,
ezhangytong_2006@163.com,
fzenghongmei666@yahoo.com.cn,
g37675767@163.com,
h*itbblzx@yahoo.com.cn, *Corresponding author
Editor: Brad Coates was editor of this paper
Received: 1 March 2010 | Accepted: 27 April 2010 | Published: 28 March 2011
Copyright: This is an open access paper. We use the Creative Commons Attribution 3.0 license that permits unrestricted use, provided that the paper is properly attributed.
ISSN: 1536-2442 | Volume 11, Number 37
Zhang Y-L, Kulye M, Yang F-S, Xiao L, Zhang Y-T, Zeng H-M, Wang J-H, Liu Z-X. 2011. Identification, characterization, and expression of a novel P450 gene encoding CYP6AE25 from the Asian corn borer, Ostrinia furnacalis. Journal of Insect Science 11:37 available online: insectscience.org/11.37
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