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Identification,Characterization, and Expression of P450 Gene Encoding CYP6BQ13v2 from the Red Flour Beetle,Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae)
引用本文:XU,Yong-qiang WANG,Jin-jun JIANG,Hong-bo DOU,Wei TANG,Pei-an AN,Feng-ming. Identification,Characterization, and Expression of P450 Gene Encoding CYP6BQ13v2 from the Red Flour Beetle,Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae)[J]. 中国农业科学(英文版), 2009, 8(10): 1210-1218. DOI: 10.1016/S1671-2927(08)60331-4
作者姓名:XU  Yong-qiang WANG  Jin-jun JIANG  Hong-bo DOU  Wei TANG  Pei-an AN  Feng-ming
作者单位:Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716,P.R. China 
基金项目:Acknowledgements We thank David R. Nelson for his assistance with the naming of CYP6BQ13v2. This research was funded in part by the National Natural Sciences Foundation of China (30871631), the Program for New Century Excellent Talents in University, China (NCET-04-0854), and the Specialized Research Fund for the Doctoral Program of Higher Education of China (200806350009).
摘    要:
An allele of CYP6BQI3, named CYP6BQ 13v2 (GenBank accession no. FJ209361), was isolated from the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) by RT-PCR. The cDNA sequence of CYP6BQ13v2, 1 563 bp in length, contains an open reading frame of 1 554 nucleotides encoding a putative protein of 518 amino acid residues with a predicted molecular weight of 59.92 kDa and a theoretical pl of 7.60. The putative protein contains the classic hemebinding sequence motif F××G×××C×G (residues 456-465) conserved among all P450 enzymes as well as other characteristic motifs of all cytochrome P450s. It shares 98% identity with the previously published sequence of CYP6BQ13 (GenBank accession no. XP967146) from the T. castaneum genome project. Phylogenetic analysis of amino acid sequences from members of various P450 families indicated that there was closer phylogenetic relationship of CYP6BQ 13v2 with CYP302A1 and CYP307A1 mediating synthesis of the insect molting hormone, distant relationship with CYP6B1 metabolizing plant allelochemicals, CYP6D 1 linking to pyrethroid resistance and other members of CYP6 family. Expression test of the gene in the adults and immature stages of T. castaneum by quantitative real-time PCR revealed that CYP6BQ13v2 is expressed in all life stages investigated. The mRNA expression level in 1st instar larvae was 14.9- and 3.86-fold higher than those in pupae and adults, respectively. The CYP6BQ13v2 expression levels appeared in the order of 1st instar larvae, followed by 4th instar larvae, 7th instar larvae, adult, and pupae from high to low. The more bioinformation of CYP6BQ 13v2 was also analyzed.

关 键 词:CYP6BQ  P450  甲虫类  植物病虫害
收稿时间:2008-12-16

Identification, Characterization, and Expression of P450 Gene Encoding CYP6BQ13v2 from the Red Flour Beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae)
XU Yong-qiang,WANG Jin-jun,JIANG Hong-bo,DOU Wei,TANG Pei-an,AN Feng-ming. Identification, Characterization, and Expression of P450 Gene Encoding CYP6BQ13v2 from the Red Flour Beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae)[J]. 《Agricultural Sciences in China》, 2009, 8(10): 1210-1218. DOI: 10.1016/S1671-2927(08)60331-4
Authors:XU Yong-qiang  WANG Jin-jun  JIANG Hong-bo  DOU Wei  TANG Pei-an  AN Feng-ming
Affiliation:Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716,P.R. China
Abstract:
An allele of CYP6BQ13, named CYP6BQ13v2 (GenBank accession no. FJ209361), was isolated from the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) by RT-PCR. The cDNA sequence of CYP6BQ13v2, 1 563 bp in length, contains an open reading frame of 1 554 nucleotides encoding a putative protein of 518 amino acid residues with a predicted molecular weight of 59.92 kDa and a theoretical pI of 7.60. The putative protein contains the classic berne-binding sequence motif F××G×××C×G (residues 456-465) conserved among all P450 enzymes as well as other characteristic motifs of all cytochrome P450s. It shares 98% identity with the previously published sequence of CYP6BQ13 (GenBank accession no. XP_967146) from the T. castaneum genome project. Phylogenetic analysis of amino acid sequences from members of various P450 families indicated that there was closer phylogenetic relationship of CYP6BQ13v2 with CYP302A1 and CYP307A1 mediating synthesis of the insect molting hormone, distant relationship with CYP6B1 metabolizing plant allelochemicals, CYP6D1 linking to pyrethroid resistance and other members of CYP6 family. Expression test of the gene in the adults and immature stages of T. castaneum by quantitative real-time PCR revealed that CYP6BQ13v2 is expressed in all life stages investigated. The mRNA expression level in 1st instar larvae was 14.9-and 3.86-fold higher than those in pupae and adults, respectively. The CYP6BQ13v2 expression levels appeared in the order of 1st instar larvae, followed by 4th instar larvae, 7th instar larvae, adult, and pupae from high to low. The more bioinformation of CYP6BQ13v2 was also analyzed.
Keywords:Tribolium castaneum  cytochrome P450  real-time PCR analysis  bioinformatics
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