中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2009年
5期
390-394
,共5页
寇宇%于新芬%叶榕%潘劲草%崔峰%乔传令
寇宇%于新芬%葉榕%潘勁草%崔峰%喬傳令
구우%우신분%협용%반경초%최봉%교전령
库蚊属%免疫表型分型%表达的序列标记%内切核酸酶类
庫蚊屬%免疫錶型分型%錶達的序列標記%內切覈痠酶類
고문속%면역표형분형%표체적서렬표기%내절핵산매류
Culex%Immunophenotyping%Expressed sequence tags%Endonucleases
目的 研究自然种群尖音库蚊复组有机磷抗性相关酯酶B2基因EstB2杂合型的分子特征.方法 采集杭州市自然种群库蚊,提取单蚊DNA基因组,采用聚合酶链反应-限制性片段长度多态性(PCR-restriction fragment length polymorphism,PCR-RFLP)方法进行抗性相关酯酶的基因分型.对酯酶B2基因Estβ2型蚊虫的PCR产物,进一步选用Bfm Ⅰ内切酶进行酶切,确定酯酶B2基因Estβ2纯合犁及Estβ2杂合型.本次研究共提取207只单蚊DNA,156份PCR样本阳性,阳性率75.36%(156/207).结果 在156份PCR样本阳性,酯酶B2基因Estβ2型占28.20%(44/156);对该基因型进一步分型,其中Estβ2纯合型占90.90%(30/33),EstB2杂合型占9%(3/33);Estβ2杂合型是以Estβ2和一种亚型杂合(Estβ2/Estβ2<'1>)形式存在.结论 建立了在自然种群尖音库蚊复组中酯酶B2基因分型方法,研究证明了杭州自然种群库蚊存在一种有机磷抗性相关酯酶B2基因Estβ2杂合型.
目的 研究自然種群尖音庫蚊複組有機燐抗性相關酯酶B2基因EstB2雜閤型的分子特徵.方法 採集杭州市自然種群庫蚊,提取單蚊DNA基因組,採用聚閤酶鏈反應-限製性片段長度多態性(PCR-restriction fragment length polymorphism,PCR-RFLP)方法進行抗性相關酯酶的基因分型.對酯酶B2基因Estβ2型蚊蟲的PCR產物,進一步選用Bfm Ⅰ內切酶進行酶切,確定酯酶B2基因Estβ2純閤犛及Estβ2雜閤型.本次研究共提取207隻單蚊DNA,156份PCR樣本暘性,暘性率75.36%(156/207).結果 在156份PCR樣本暘性,酯酶B2基因Estβ2型佔28.20%(44/156);對該基因型進一步分型,其中Estβ2純閤型佔90.90%(30/33),EstB2雜閤型佔9%(3/33);Estβ2雜閤型是以Estβ2和一種亞型雜閤(Estβ2/Estβ2<'1>)形式存在.結論 建立瞭在自然種群尖音庫蚊複組中酯酶B2基因分型方法,研究證明瞭杭州自然種群庫蚊存在一種有機燐抗性相關酯酶B2基因Estβ2雜閤型.
목적 연구자연충군첨음고문복조유궤린항성상관지매B2기인EstB2잡합형적분자특정.방법 채집항주시자연충군고문,제취단문DNA기인조,채용취합매련반응-한제성편단장도다태성(PCR-restriction fragment length polymorphism,PCR-RFLP)방법진행항성상관지매적기인분형.대지매B2기인Estβ2형문충적PCR산물,진일보선용Bfm Ⅰ내절매진행매절,학정지매B2기인Estβ2순합리급Estβ2잡합형.본차연구공제취207지단문DNA,156빈PCR양본양성,양성솔75.36%(156/207).결과 재156빈PCR양본양성,지매B2기인Estβ2형점28.20%(44/156);대해기인형진일보분형,기중Estβ2순합형점90.90%(30/33),EstB2잡합형점9%(3/33);Estβ2잡합형시이Estβ2화일충아형잡합(Estβ2/Estβ2<'1>)형식존재.결론 건립료재자연충군첨음고문복조중지매B2기인분형방법,연구증명료항주자연충군고문존재일충유궤린항성상관지매B2기인Estβ2잡합형.
Objective To investigate the heterozygous genotype and molecular characteristics of Organophosphorus resistance associated with heterozygous Estβ2 of esterase B2 gene from natural population of Culex pipiens complex. Methods Genomic DNA was extracted from natural populations of Culex pipiens complex in Hangzhou. The PCR-restriction fragment length polymorphism (PCR-RFLP) assay was applied to type the resistance associated esterase gene. Estβ2 of esterase B2 gene was identified by PCR-RFLP, and the genotyping for heterozygous Estβ2 was carried out after restriction enzyme digesting by Bfm I endonuclease. Results The DNA was isolated from 207 Culex pipiens respectively,while 156 PCR samples showed positive and the positive rate was 75.36% (156/207). The PCR-RFLP assay of esterase B2 gene revealed that the Estβ2 was accounted about 28.20% (44/156) in 156 positive samples. There were two genotypes identified, namely homozygous Est,2 (90. 90%, 30/33) and heterozygous Estβ2 (9%, 3/33), heterozygous Estβ2 was in existence of a hybrid form as which combined with Estβ2 and a subtype(Estβ2/ Estβ2<'1>). Conclusion Heterozygous Estβ2 of Organophosphorus resistance associated with esterase genotype was determined in natural population of Culex pipiens, and a genotyping method was established.
