中国媒介生物学及控制杂志

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复合PCR-RFLP技术用于尖音库蚊复组抗性相关酯酶基因分子分型

寇宇1;叶榕1;潘劲草1;乔传令2;崔峰2;于新芬1;黄志成1   

  1. 1杭州市疾病预防控制中心媒介生物控制与消毒所 杭州310006;2中国科学院动物研究所农业虫鼠害综合治理研究国家重点实验室
  • 出版日期:2007-08-20 发布日期:2007-08-20

Molecular typing on insecticide resistance-associated esterase genes of Culex pipiens complex by PCR-RFLP techniques

KOU Yu;YE Rong; PAN Jin-cao; QIAO Chuan-ling; CUI Feng; YU Xin-fen; HUANG Zhi-cheng   

  1. Hangzhou Municipal Center for Disease Control and Prevention, Hangzhou 310006, China
  • Online:2007-08-20 Published:2007-08-20

摘要: 目的 应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)检测法,建立尖音库蚊抗性酯酶等位基因分型方法。方法 根据尖音库蚊抗性酯酶基因保守区序列设计引物;提取单个蚊虫基因组DNA,采用PCR-RFLP分析的方法,对抗性酯酶等位基因分型。结果 酯酶基因扩增片段长度为860 bp,经限制性内切酶酶切后,尖音库蚊抗性Est-β11纯合型有5条带(70、89、147、198和347 bp);Est-β2纯合型出现4条带(60、147、207和437 bp);Est-β11/Est-β2杂合型出现5条带;Est-βN纯合型出现3条带(147、207和506 bp)。结论 PCR-RFLP方法能快速、有效地检测尖音库蚊抗性酯酶等位基因类型。

关键词: 尖音库蚊复组, 抗性基因, 聚合酶链反应-限制性片段长度多态性, 分子分型

Abstract: Objective Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) assay was applied to molecular typing of resistance-associated esterase genes of Culex pipiens complex.Methods The primers specific to the conserved region of resistance-associated esterase genes of C.pipiens were designed.The genomic DNA was extracted from single mosquito.PCR-RFLP were used in molecular typing of resistance-associated esterase genes.Results Csp6Ⅰ digestion pattern of five fragments(70 bp,89 bp,147 bp,198 bp,347 bp) was noted for homozygous resistance-associated esterase-β11,and the pattern of four fragments(60 bp,147 bp,207 bp,437 bp) was noted for homozygous resistance-associated esterase-β2,and the patterm of five fragments was noted for heterozygous Est-β11/Est-β2,and the pattern of three fragments(147 bp,207 bp,506 bp) was noted for homozygous Est-βN.Conclusion PCR-RFLP is an effective,specific method for molecular typing of resistance-associated esterase genes of C.pipiens complex.