应用微生物发酵技术获得马铃薯甲虫脯氨酸脱氢酶基因双链RNA的初步研究  

Preliminary Study on Techniques for Bacterial Expression of Dsrna from Proline Hydrogenase Gene in Leptinotarsa decemLineata(Say)

作　　者：吕东[1] 杨静[1] 李国清[1] 

机构地区：[1]南京农业大学昆虫系农业部病虫害监测与治理重点开放实验室,南京210095

出　　处：《农药》2010年第8期571-573,共3页Agrochemicals

基　　金：公益性行业科研专项基金项目(200803024)

摘　　要：RNA干扰技术在植物保护领域具有潜在的应用前景,寻找经济、实用的生产技术十分必要。将外源碱基序列通过载体转入基因工程菌,发酵增殖而获得大量目标dsRNA的方法是备选技术之一。建立了利用大肠杆菌表达目的基因双链RNA的方法,通过改变诱导子IPTG的浓度及诱导时间优化了发酵条件,初步肯定了此项技术应用于药剂开发的可能性。RNA interference shows great promise for use in insect pest control.It is necessary to develop safe and effective techniques to produce large quantities of dsRNA.A possible way is to apply engineered bacterium to produce a massive amount of dsRNA in vivo.In the present paper,a dsRNA-produced vector was constructed and transferred to Escherichia coli HT115.The engineered bacterium was cultured,and a great amount of dsRNA derived from proline hydrogenase gene in L.decemlineata was obtained.Moreover,the fermentation condition was optimized in order to get higher expression level of dsRNA.This is the first step to develop novel insecticides by RNA interference techniques.

关 键 词：RNA干扰 dsRNA表达载体 大肠杆菌HT115 新型农药 

分 类 号：TQ453.2[化学工程—农药化工]