白纹伊蚊基因组中CYP6家族新成员基因片段的克隆与分析  

CLONE AND ANALYSIS ON GENE FRAGMENTS OF NEW MEMBERS OF CYP6 FAMILY FROM GENOME OF AEDES ALBOPICTUS

作　　者：吴瑜[1] 黄炯烈[1] 周国理[1] 柳晓欣[1] 钟作良[1] 

机构地区：[1]中山医科大学寄生虫学教研室,广州510080

出　　处：《寄生虫与医学昆虫学报》2001年第1期23-30,共8页Acta Parasitologica et Medica Entomologica Sinica

基　　金：广东省自然科学基金!编号 :970 0 92 ;"2 11工程"重点学科建设基金资助!编号 :98124

摘　　要：目的 :获得白纹伊蚊 CYP6基因家族中新成员的基因片段。方法 :根据家蝇 CYP6 A1、 CYP6 D1以及致倦库蚊 CYP6 E1同源区氨基酸序列 ,设计 1对简并引物 ,分别以白纹伊蚊敏感品系和抗性品系基因组 DNA为模板进行 PCR,产物经 T- A克隆转化至 JM10 9大肠杆菌中 ,经筛选后测序 ,获得与设计的细胞色素 P4 50基因片段大小相符合的 5个基因片段 ,并将推导的氨基酸序列进行同源性分析。结果 :在白纹伊蚊敏感品系中获得了 2个基因片段 ( A EDG- S1,AEDG- S2 ) ,核苷酸序列长度分别为 2 4 0 bp和 2 36 bp;在白纹伊蚊溴氰菊酯抗性品系中获得了 3个基因片段 ( A EDG- R2 ,A EDG- R3,AEDG- R4 ) ,核苷酸序列长度分别为 2 36 bp、 2 36 bp和 2 39bp。所得序列与昆虫 CYP6家族的同源性在 2 3.1%～ 53.8%之间 ,与 CYP3家族的同源性在 2 5.6 %～4 3.9%之间 ,而与 CYP4家族同源性较低 ,为 13.9%～ 2 4 .4 %。结论 :所得 5个序列为 CYP6家族中 5个新成员的基因片段。Aim:To obtain new members'gene fragments of CYP6 family from Aedes albopictus. Method: A pair of degenerate primers according to the homologous amino acid sequences of CYP6A1,CYP6D1 in Musca domestica and CYP6E1 in Culex quinquefasciatus was designed and used to amplify CYP6 gene from genomic DNA of sensitive and deltamethrin resistant strains of Ae.albopictus by PCR technique. Five DNA fragments sized as designed were obtained. These fragments were recombined by T A ligation, and cloned into JM109.Five positive clones were selected to sequence. Homologous analysis on these deduced amino acid sequences were executed. Results: Five nucleotide sequences were obtained, of which the length is from 236bp to 240bp.The identities of these cloned sequences to other known members of CYP6 are from 23 1% to 53 8% ,to CYP3 are from 25 6% to 43 9% ,and to CYP4 are from 13 9% to 24 4% . Conclusion:These cloned sequences are gene fragments from new members of CYP6 family.

关 键 词：白纹伊蚊 基因组 CYP6 同源性分析 杀虫剂 抗性 

分 类 号：R184[医药卫生—流行病学]