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Issue:ISSN 1000-7083
          CN 51-1193/Q
Director:Sichuan Association for Science and Technology
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Your Position :Home->Past Journals Catalog->2016 Vol.35 No.3

Cloning and Expression of Cytochrome P450 CYP6J1 from Aphis gossypii
Author of the article:WANG Yamei1, HUANG Lina1, AI Xinyu1, WEI Yuanjie1, LIU Xiaoning1*, GAO Xiwu2*
Author's Workplace:1. College of Life Sciences and Technology, Xinjiang University, Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, Urumqi 830046, China;
2. College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China
Key Words:Aphis gossypii; P450 CYP6J1; cloning; protein expression
Abstract:In order to understand the structure and function of cytochrome P450 CYP6J1 protein, P450CYP6J1 gene from cotton aphid (Aphis gossypii) was cloned and analyzed using bioinformatic method. P450 CYP6J1 protein was first expressed in prokaryotic, followed by SDS-PAGE and Western-blot detection. The results showed that the length of open reading frame region was 1398 bp, encoding 465 amino acids residues with the predicted molecular weight of 53.67 kD and isoelectric point of 8.80. Amino acid sequence analysis showed that P450 CYP6J1 protein had no signal peptide. Homology analysis showed that the amino acids of P450 CYP6J1 of A. gossypii was similar with that of pea aphid (Acyrthosiphon pisum) (homologous similarity 92%). The His-CYP6J1 protein was obtained after being expressed in Escherichia coli BL21, and the target protein was proved by Western-blot. These results provided a foundation for preparation of P450 CYP6J1 polyclonal antibody from A. gossypii.
2016,(35): 378-383 收稿日期:2015-12-08
贺丽虹, 赵淑娟, 胡之璧. 2008. 植物细胞色素P450基因与功能研究进展[J]. 药物生物技术, 15(2):142-147.
邱星辉, 何凤琴, 李梅. 2003. 细胞色素P450介导的昆虫抗药性[J]. 生命的化学, 23(4):279-281.
张学涛, 柳建伟, 李芬, 等. 2012. 北疆地区棉蚜对不同杀虫剂敏感度水平测定[J]. 植物保护, 38(2):163-166.
Ahuja I, Rohloff J, Bones AM. 2010. Defence mechanisms of Brassicaceae:implications for plant-insect interactions and potential for integrated pest management. A review[J]. Agronomy for Sustainable Development, 30(2):311-348.
Amenya DA, Naguran R, Lo TC, et al. 2008. Over expression of a cytochrom P450 (CYP6P9) in major African malaria vector, Anopheles funestus, resistant to pyrethroids[J]. Insect Biochemistry and Molecular Biology, 17(1):19-25.
Carino FA, Koener JF, Plapp Jr. FW, et al. 1994. Constitutive overexpression of the cytochrome P450 gene CYP6A1 in a house fly strain with metabolic resistance to insecticides[J]. Insect Biochemistry and Molecular Biology, 24(4):411-418.
Crinnion WJ. 2009. Chlorinated pesticides:threats to health and importance of detection[J]. Alternative Medicine Review, 14:347-359.
Deeni YY, Paine MJ, Ayrton AD, et al. 2001. Expression, purification, and biochemical characterization of a human cytochrome P450 CYP2D6-NADPH cytochrome P450 reductase fusion protein[J]. Archives of Biochemistry and Biophysics, 396(1):16-24.
Denholm I, Rowland MW. 1992. Tactics for managing pesticide resistance in arthropods:theory and practice[J]. Annual Review of Entomology, 37(1):91-112.
Eldefrawi ME, Miskus R, Sutcher V. 1960. Methylenedioxyphenyl derivatives as synergists for carbamate insecticides on susceptible, DDT and parathion-resistant house flies[J]. Economic Entomology, (53):231-234.
Feyereisen R, Andersen JF, Carino FA, et al. 1995. Cytochrome-P450 in the house-fly-structure, catalyticactivity and regulation of expression of Cyp6a1 in an insecticide-resistant strain[J]. Pesticide Science, 43:233-239.
Graslund S, Nordlund P, Weigelt J, et al. 2008. Protein production and purification[J]. Nature Methods, 5(2):135-146.
Komarek M, Cadkova E, Chrastny V, et al. 2010. Contamination of vineyard soils with fungicides:a review of environmental and toxicological aspects[J]. Environment International, 36:138-151.
Kubota A, Kim EY, Jwata H. 2009. Alkoxyresorufin (methoxy-, ethoxy-, pentoxy- and bexzyloxyresorufin 0-dealkylase activities by in vitro-expressed cytochrome P450 1A4 and 1A5 from common cormorant (Phalacrocorax carbo)[J]. Comparative Biochemistry and Physiology, Part C, 149:544-551.
Liu XN, Zhang L, Zhang XT, et al. 2013. Molecular cloning and recombinant expression of cytochrome P450 CYP6B6 from Helicoverpa armigera in Escherichia coli[J]. Molecular Biology Reports, 40:1211-1217.
Scott JG, Lee SST. 1993. Purification and characterization of a cytochrome P450 from insecticide susceptible and resistant strains of housefly, Musca domestica L., before and after phenobarbital exposure[J]. Archives of Insect Biochemistry and Physiology, 24(1):1-19.
Scott JG, Wen Z. 2001. Cytochromes P450 of insects:the tip of the iceberg[J]. Pest Management Science, 57(10):958-967.
Snyder MJ, Van Antwerpen. 1998. Evidence for a diazepam-binding inhibitor (DBI) benzodiazepine receptor-like mechanism in ecdysteroidogenesis by the insect prothoracic gland[J]. Cell Tissue Research, 294:161-168.
Tan W,Guo Y. 1996. Effects of host plant on susceptibility to deltamethrin and detoxification enzyme of Heliothis armigera (Lepidoptera:Noctuidea)[J]. Economic Entomology, 86:11-14.
Uchinomiya S, Nonaka H, Wakayama S, et al. 2013. In-cell covalent labeling of reactive His-tag fused proteins[J]. Chemical Communications, 49:5022-5024.
Wang XP, Hobbs AA. 1995. Isolation and sequence analysis of a cDNA clone for a pyrethroidinducible cytochrome P450 from Helicoverpa armigera[J]. Insect Biochemisry and Molecular Biology, 25(9):1001-1009.
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