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[目的 ]探讨淡色库蚊细胞色素P45 0与溴氰菊酯抗药性的关系。 [方法 ]采用一对昆虫细胞色素P45 0简并引物 ,以反转录 聚合酶链反应从淡色库蚊扩增特异片段 ,T/A直接克隆法筛选阳性克隆 ,对新序列进行cDNA芯片和逆Northern分析。 [结果 ]从淡色库蚊对溴氰菊酯敏感品系和抗性品系获得 112个阳性克隆 ,其中 2 4个阳性克隆测序后显示为细胞色素P45 0新序列 ,由GenBank登录上网 ;经国际细胞色素P45 0命名委员会鉴定分别属CYP4家族CYP4C、CYP4D、CYP4H和CYP4J等 4个亚家族 ;2 4个CYP4cDNA片段中 ,来自敏感品系的 2个片段(NYDS3和NYDS5 )和来自抗性品系的 4个片段 (NYDR6、NYDR9、NYDR15和NYDR17)在两品系间存在差别 ,cDNA芯片信号亮度值均是抗性品系大于敏感品系 ,倍数在 3 .1～ 9.7范围内 ;NYDR17仅与抗性探针杂交 ;逆Northern再鉴定 ,获得了与cDNA芯片一致的结果。 [结论 ]淡色库蚊CYP4与溴氰菊酯抗性相关 ;在淡色库蚊溴氰菊酯抗性机理中 ,可能存在P45 0基因点突变而导致的特异表达。
[Objective] The research aimed to discuss the relationship between cytochrome P450 of Culex pipiens pallens and resistance to deltamethrin. [Method] A pair of insect cytochrome P45 0 degenerate primers were used to amplify specific fragments from Culex pipiens pallens by reverse transcription polymerase chain reaction. The positive clones were screened by T / A direct cloning method, Northern analysis. [Result] 112 positive clones were obtained from Culex pipiens pallens susceptible and susceptible strains of deltamethrin, of which 24 positive clones were sequenced and displayed as a new sequence of cytochrome P45 0, which was registered online by GenBank. The P45 0 nomenclature committee identified 4 subfamilies of the CYP4 family, CYP4C, CYP4D, CYP4H and CYP4J, respectively; of the 24 CYP4cDNA fragments, two fragments from the susceptible strain (NYDS3 and NYDS5) and four fragments from the resistant strain (NYDR6, NYDR9, NYDR15 and NYDR17) were different between the two lines. The signal intensity of the cDNA chips were all higher than that of the susceptible lines, the multiple of which ranged from 3.1 to 9.7. The NYDR17 only hybridized with the resistant probe. The reverse Northern re-identification, obtained consistent with the cDNA chip results. [Conclusion] CYP4 of Culex pipiens pallens was related to resistance to deltamethrin. Among deltamethrin resistance mechanisms of Culex pipiens pallens there may be specific expression caused by point mutation of P45 0 gene.