Overexpression of cytochrome P450 genes in pyrethroid-resistant Culex quinquefasciatus
Overexpression of cytochrome P450 genes in pyrethroid-resistant Culex quinquefasciatus
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DOI：
10.1016/j.ibmb.2010.01.006
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发表时间：
2010-02-01
影响因子：
3.8
通讯作者：
Tomita, Takashi
中科院分区：
文献类型：
--
作者：
Komagata, Osamu;Kasai, Shinji;Tomita, Takashi
致倦库蚊幼虫对拟除虫菊酯类杀虫剂具有极高的抗性，而成虫对拟除虫菊酯类杀虫剂的抗性较低。微体单加氧酶代谢增加是该菌株抗性的主要因素之一。我们从JPal-per菌株中克隆了46个新的细胞色素P450 cdna。我们设计了一个寡核苷酸微阵列，用于新的46个基因和16个先前报道的P450基因以及其他非P450基因探针。其中，5个P450基因在JPal-per幼虫中与敏感菌株相比上调（>2.5倍）。在芯片分析中筛选到的5个P450基因中，CYP9M10、CYP4H34和CYP6Z10的表达率最高，qPCR进一步验证了这3个基因的表达率分别为264倍、8.3倍和3.9倍。在JPal-per中，CYPSIM10和C 'YP4H34的转录水平表现出类似的阶段依赖模式，在日本小asawara群岛的幼虫中，CYPSIM10和C 'YP4H34的转录水平在日本小asawara群岛的幼虫中处于高表达水平，在成虫期急剧下降。这两个基因的幼虫特异性过表达方式与先前报道的JPal-per菌株的阶段依赖性抗性特征相一致，表明这两个p450基因CYP9M10和CYP4H34参与了JPal-per菌株的拟除虫菊酯类脱毒。(C) 2010 Elsevier Ltd.版权所有。
JPal-per strain of Culex quinquefasciatus exhibits extremely high resistance against pyrethroids in larvae, though the resistance is greatly lower in adults. Increased microsome monooxygenase metabolism is one of the major factors of the larval resistance in this strain. We cloned 46 novel cytochrome P450 cDNAs from JPal-per strain. An oligonuclecitide microarray was designed for the novel 46 genes plus 16 previously reported P450 genes along with other non-P450 gene probes. Of these, five P450 genes were upregulated (>2.5-fold) in JPal-per larvae as compared with a susceptible strain. The expression ratios for the highest three among the five P450 genes screened in the microarray analysis, CYP9M10, CYP4H34 and CYP6Z10, were further validated by qPCR as 264-, 8.3-, and 3.9-fold, respectively. In JPal-per, the transcription levels of CYPSIM10 and C`YP4H34 showed a similar stage-dependent pattern as a high expression level during the larvfrom Ogasawara Islands in Japanal stage dramatically decreases in the adult stage. This larval specific overexpression manner of the two genes was consistent with the characteristic of stage-dependent resistance of JPal-per strain previously reported, suggesting that the two P450s, CYP9M10 and CYP4H34, are involved in pyrethroid detoxification in JPal-per strain. (C) 2010 Elsevier Ltd. All rights reserved.