蚊虫RNAi显微注射方法的建立  

MICROINJECTION METHOD FOR RNAI ON ADULT MOSQUITOES

作　　者：吴群[1,2] 李春晓[2] 廖承红[1] 韩谦[1] 赵彤言[2] 

机构地区：[1]海南大学农学院,海口570228 [2]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071

出　　处：《寄生虫与医学昆虫学报》2015年第4期233-237,共5页Acta Parasitologica et Medica Entomologica Sinica

基　　金：国家重大专项课题（No.2012zx10004-219）和国家自然科学基金（81371847）

摘　　要：本文构建了蚊虫显微注射方法.以致倦库蚊Culex pipiens quinquefasciatus成蚊为注射对象,从不同麻醉方法对蚊虫进行固定、注射蚊虫身体不同部位、注射剂量等方面进行优化来构建最佳显微注射方案.结果显示蚊虫冰冻（-30℃）4、5、7、10 min后蚊虫成活率分别为98.97％、89.63％、83.50％、79.87％,4 min麻醉效果最佳;对蚊虫充CO2气体持续0.5、1、2、4 min后蚊虫成活率均为100％;注射剂量为0.7～0.8 μL时可以明显看出蚊虫腹部变大;正对胸部腹面注射蚊虫的成活率67.81％,低于从胸部侧面注射成活率93.85％.结果表明选取羽化3～5d并断糖水4h蚊虫于-30℃冰冻4 min后放置于冰盒上,利用微量注射器从蚊虫胸部侧面注入0.7 ～0.8 μL dsRNA溶液是最佳的注射方法..In the study, microinjection method for RNAi was established in Culex pipiens quinquefasciatus. To obtain successful injection, mosquito fixation, injection site and injection volume was optimized. After being frozen at-30℃ for 4, 5, 7 and 10 min, the survival rate of mosquitoes was 98.97% , 89. 63% , 83.50% and 79. 87% respectively. Obviously, four minutes was suitable for injection in view of movements and freezing damages. Furthermore, the experiments indicated that the ideal injection volume was 0. 7 - 0. 8 μL and the best injection site was lateral thorax with survival rate of 94%.

关 键 词：微量注射 致倦库蚊 成蚊 

分 类 号：Q969.442.1[生物学—昆虫学]