【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。更多还原

【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.更多还原