mRNA差异显示法筛选和克隆东亚飞蝗抗药性相关基因  

Screening and Cloning of the Oriental Migratory Locust-related Resistance Gene by mRNA Differential Display

作　　者：陶杰[1] 单颖[1] 荆迎军[1] 吕宪禹[1] 李德森[1] 谢建军 张杰 

机构地区：[1]南开大学生命科学学院,天津300071 [2]天津市防蝗指挥部,天津300380

出　　处：《南开大学学报（自然科学版）》2009年第1期107-111,共5页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：天津市财政局农林处基金(03360491)

摘　　要：筛选和克隆东亚飞蝗抗药性相关基因.采用mRNA差异显示法(mRNA differential display PCR, DD-PCR),对比了东亚飞蝗在受到氯氰菊酯杀虫剂的诱导前后,基因表达发生的变化.对差异片段进行了回收,克隆、测序,Northern点杂交验证以及序列分析和同源性比较.东亚飞蝗在受到氯氰菊酯杀虫剂的诱导前后,存在明显的基因表达差异,发现差异表达条带9条,对9条差异表达条带进行克隆、测序,经序列分析和同源性比较和RNA点杂交验证,表明确认其中1条带与东亚飞蝗的抗药性有关.东亚飞蝗的mRNA差异显示证明,该EST序列可能在抗药的发生发展中具有一定的作用.To isolate and clone the oriental migratory locust-related resistance gene. The specially expressed cDNA fragments of the oriental migratory locust-related resistance were screened by differential display polymerase chain reaction （DD-PCR） after extracting the total RNA from tissues of the oriental migratory locust which were infected with cypermethrin and normal. Then these fragments were cloned to plasmid pBS-T and subsequently underwent se- quence analysis, homogenous comparing and RNA dot hybridization. Obvious difference in gene expression between the oriental migratory locust were infected with cypermethrin and norreal oriental migratory locust were found in 9 fragments express. Among them 9 fragments were cloned and by sequence analysis, homogenous comparing and RNA dot hybridization, one of them showed the oriental migratory locust-related resistance gene. The specially expressed EST sequence might be the oriental migratory locust-related resistance gene and may be play a certain role in the development of drug resistance.

关 键 词：东亚飞蝗幼虫 诱导 MRNA差异显示 昆虫抗药性的系统 BLAST 

分 类 号：Q78[生物学—分子生物学]