溴氰菊酯胁迫下小菜蛾差异表达基因的筛选与分析  

Screening and Analysis of the Differential Expressed Sequence of Diamondback Moth,Plutella xylostella

作　　者：刘白朵[1] 李风良[2] 和碧蕾[1] 于希忠[1] 李忠英[2] 程罗根[1] 

机构地区：[1]南京师范大学生命科学学院,江苏南京210046 [2]贵州省农业科学院植物保护研究所,贵州贵阳550006

出　　处：《南京师大学报（自然科学版）》2010年第2期91-95,共5页Journal of Nanjing Normal University(Natural Science Edition)

基　　金：国家自然科学基金(30560085);南京师范大学实践创新训练计划项目

摘　　要：通过双向cDNA RDA,即分别以敏感品系为检测子(tester)、抗溴氰菊酯品系为驱赶子(driver)和以抗溴氰菊酯品系为检测子(tester)、敏感品系为驱赶子(driver)筛选2种品系中的差异表达基因,初步分析小菜蛾敏感品系和抗性品系之间的遗传差异.经过3轮消减杂交后,将所得的差异片段克隆于PMD 18-T载体,氨苄筛选阳性克隆,经菌落PCR鉴定后送样测序.得到2条序列与GenBank数据库中的部分已知序列有一定同源性,其中1条序列与编码S3a蛋白基因有较高的同源性.We sifted the differential expressed sequence from Plutella xylostella susceptible strain and the deltamethrin-resistance strain by two-way cDNA representational difference analysis（cDNA RDA）.The two-way cDNA RDA was performed respectively using cDNA from the P.xylostella susceptible strain as Tester amplicon and from the deltamethrin-resistance strain as Driver amplicon,and using cDNA from the deltamethrin-resistance strain as Tester amplicon and from the susceptible strain as Driver amplicon.The differential sequences between Plutella xylostella susceptible strain and the deltamethrin-resistance strain were analysed preliminarily.After three rounds of subtractive hybridization,the differential fragments were cloned with pMD18-T vector,then screening positive clone with ampicillin and sequenced after colony PCR.Comparisons between our sequencing result and data in GenBank showed that there were two sequences appearing relatively high homology to some known sequences.One of them had high homology with the sequence encoding s3a protein.

关 键 词：小菜蛾 抗性遗传 菌落PCR CDNA RDA 

分 类 号：N[自然科学总论]