目的构建淡色库蚊糖原分支酶基因(GBE)的昆虫表达载体,通过细胞转染法鉴定其与蚊虫抗药性相关的关系。方法用基因重组方法构建GBE的昆虫表达载体;通过脂质体转染法将昆虫表达载体转入伊蚊C6/36细胞,继而通过观察细胞形态及3H TdR掺入法检测稳定表达GBE基因的细胞对溴氰菊酯杀虫剂的敏感性。结果成功构建GBE基因的昆虫表达载体同时通过筛选获得稳定表达该基因的细胞株。3H TdR掺入法结果显示转染GBE基因能够提高细胞的存活率,并与对照组差异有显著性(P〈0.05)。结论 GBE基因在与淡色库蚊溴氰菊酯抗性相关,且其在细胞对抗溴氰菊酯的毒性作用中有保护作用。
[ Objective ] To construct prokaryotic and insect expression plasmids of glycogen-branching enzyme (GBE) and further research the relationship between NYD-GBE and insecticide resistance. [ Methods ] The insect expression plasmids of GBE were constructed by gene re-construction methods. The recombinant insect expression vectors were transfected into Aedes albopictus C6/36 cells. The viability of the transfected cells and untransfected ones were detected by 3H-TdR. [ Results ] The expressed plasmids pIB/VS-His/GBE were re-constructed successfully. Then it was confirmed that the expression veetors of insect cells were successfully constructed and the stabletransfeeted cell lines were established. We found that there was significant difference compared with the control (P 〈 0.05). [ Conclusion ] GBE gene has association with the dehamethrin resistance of Cx. pipiens pallens.