Molecular mechanism of acetylcholinesterase insensitivity to insecticide and its utilization.

乙酰胆碱酯酶对杀虫剂不敏感的分子机制及其应用。

• 批准号: 14206005
• 负责人: KONO Yoshiaki
• 金额: $ 31.37万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14206005/
• 关键词: acetylcholinesterase; insecticide resistance; insensitivity to insecticide; amino acid substitution; gene expression; purification of cnzyme; purification of enzyme; organophosphate; 酸素精製; コガタアカイエカ; 分子機構; アブラムシ; ハダニ; 殺虫剤感受性; cDNA

Molecular mechanism of acetylcholinesterase (AChE) insensitivity to organophosphates and carbamates was studied, and the insensitivity was elucidated to be caused by amino acid substitutions in the active cite of the enzyme. In cyclorrhapha dipterans, only one gene of AChE exists in their genome, and several amino acids were replaced in the insensitive AChE. In the orthologous AChE of other insects, however, no amino acid substitution has been found in relation to the insensitivity. After the discovery of the second AChE (Ace paralogous AChE, AP-AChE) gene in an aphid and a mosquito species in 2002, we cloned AP-AChE cDNA from several species of resistant insects and mites ; Myzus persicae, Aphis gossypii, Nephotettix cincticeps, Tetranychus urticae and Tetranychus kanzawai, and found species specific amino acid substitutions causing the insensitivity at the important position in the active site of the enzyme. When 6 recombinant AChEs having respective amino acid substitutions were expressed in baculovirus-insect cultured cell system, AChEs produced showed specific characteristics in the reduction of sensitivity to insecticides.Purification of AChE and gene expression of two AChE were also studied in the mosquitoes.

研究了乙酰胆碱酯酶(AChE)对有机磷和氨基甲酸酯类化合物不敏感的分子机制，认为这种不敏感是由于酶活性中心的氨基酸取代所致。在环纹夜蛾中，只有一个AChE基因存在于它们的基因组中，并且在不敏感的AChE中替换了几个氨基酸。然而，在其他昆虫的同源AChE中，没有发现与不敏感有关的氨基酸替代。Ace paralogous AChE(Ace paralogous AChE，AP-AChE)基因于2002年在一种蚜虫和一种蚊子中被发现后，我们从几种抗性昆虫和螨类中克隆了AP-AChE基因，并在酶活性部位的重要位置发现了物种特有的氨基酸替换导致该酶活性部位的不敏感性。在杆状病毒-昆虫培养细胞系统中表达6种氨基酸取代的重组AChE后，所产生的AChE在降低杀虫剂敏感性方面表现出独特的特性。

项目成果

Amino acid substitution in Ace paralogous acetylcholinesterase accompanied by organophosphate resistance in the spider mite Tetranychus kanzawai

• DOI: 10.1016/j.pestbp.2005.02.004
• 发表时间: 2005-06-01
• 期刊: PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
• 影响因子: 4.7
• 作者: Aiki, Y;Kozaki, T;Kono, Y
• 通讯作者: Kono, Y

Two amino acid substitutions in acetylcholinesterase associated with pirimicarb and organophosphorous insecticide resistance in the cotton aphid, Aphis gossypii Glover.

乙酰胆碱酯酶中的两个氨基酸取代与棉蚜 Aphis gossypii Glover 的抗蚜威和有机磷杀虫剂抗性相关。

• 发表时间: 2004
• 期刊: Insect Molecular Biology 13
• 作者: S.Toda;S.Komazaki;T.Tomita;Y.Kono
• 通讯作者: Y.Kono

Toshinori Kozaki, Takashi Tomita, Kiyoko Taniai, Minoru Yamakawa, Yoshiaki Kono: "Expression of two acetylcholinesterase genes from organophosphate sensitive-and insensitive-houseflies, Musca domestica L., using a baculovirus insect cell system"Applied En

Toshinori Kozaki、Takashi Tomita、Kiyoko Taniai、Minoru Yamakawa、Yoshiaki Kono：“使用杆状病毒昆虫细胞系统从有机磷敏感和不敏感的家蝇 Musca Domestica L. 中表达两种乙酰胆碱酯酶基因”Applied En

Sequence of a cDNA encoding acetylcholinesterase from susceptible and resistant two-spotted spider mite, Tetranychs urticae.

编码来自易感和抗性二斑叶螨（Tetranychs urticae）的乙酰胆碱酯酶的 cDNA 序列。

• 发表时间: 2003
• 期刊: Insect Biochem.Mol.Biol. 33
• 作者: Y.Anazawa;T.Tomita;Y.Aiki;T.Kozaki;Y.Kono
• 通讯作者: Y.Kono

cDNA and deduced protein sequence of acetylcholinesterase from the diamondback moth, Plutella xylostella(L.).

小菜蛾 Plutella xylostella(L.) 乙酰胆碱酯酶的 cDNA 和推导的蛋白质序列。

• 发表时间: 2003
• 期刊: Appl.Ent.Zool. 38
• 作者: X-Y.Ni;T.Tomita;S.Kasai;Y.Kono
• 通讯作者: Y.Kono