小菜蛾对溴氰菊酯抗性相关的DNA片段的克隆与分析  

Cloning and analysis of DNA sequences related to deltamethrin resistance in the diamondback moth, Plutella xylostella (L.)

作　　者：程罗根[1] 陈之浩[2] 张晓飞[1] 李忠英[2] 

机构地区：[1]南京师范大学生命科学学院,南京210097 [2]贵州省农业科学院植物保护研究所,贵阳550006

出　　处：《昆虫学报》2004年第4期526-529,共4页Acta Entomologica Sinica

基　　金：国家自然科学基金项目 ( 3 0 0 60 0 5 1)

摘　　要：采用代表性差异分析 (representationaldifferenceanalysis ,RDA)方法 ,以小菜蛾Plutellaxylostella (L .)的敏感品系作为对照组 ,溴氰菊酯抗性近等基因系 (nearisogenicline)作为测试组 ,通过三轮消减杂交得到大小为 15 0～ 30 0bp的差异扩增带 ,经亚克隆测序并与GenBank等数据库同源比较 ,发现差异片段与已知抗性相关基因无同源性 ;以随机选取的差异片段作探针进行Southernblot分析 ,在测试组扩增子和三轮差异产物中都获得了阳性结果 ,而在对照组扩增子中的结果是阴性的。Using representational difference analysis (RDA) technique,the genomic difference between the susceptible strain and the deltamethrin resistance near isogenic line of the diamondback moth, Plutella xylostella (L.) was preliminarily analyzed. In RDA system, the driver amplicon is from the susceptible strain while the tester amplicon from the deltamethrin resistance near isogenic line strain. After three rounds of subtractive hybridization, the differential amplification products were obtained and their sizes were about 150-300 bp. Comparisons between our sub_clone sequencing results and those recorded in GenBank show that these products have partial gene homology with Homo sapiens, bronchitis virus and so on, but show no homology to any known resistant gene. Preliminary judgment revealed that these segments might be new gene sequences or controlling gene sequences.

关 键 词：小菜蛾 溴氰菊酯 抗药性 代表性差异分析法 DNA测序 

分 类 号：Q966[生物学—昆虫学]