鸡柔嫩艾美耳球虫体内瞬时转染系统的建立  

The development of in vivo transient transfection system of Eimria tenella using yellow fluorescent protein gene as a marker

作　　者：窦刚[1] 闫文朝[2] 刘贤勇 殷光文[1] 索勋[1] 

机构地区：[1]中国农业大学动物医学院,北京海淀100193 [2]河南科技大学动物科技学院,河南洛阳471003

出　　处：《中国兽医杂志》2009年第7期17-19,共3页Chinese Journal of Veterinary Medicine

基　　金：国家"863"计划专题项目(2006AA02Z458)

摘　　要：本研究通过限制性内切酶介导的整合转染,将球虫表达载体pEtHEA转染的柔嫩艾美耳球虫子孢子经泄殖腔接种到4日龄AA雏鸡的回盲口。在不同时间段在荧光显微镜下检测黏膜内容物内虫体是否表达黄色荧光蛋白;另外还收集粪便中的卵囊,观察其是否同样表达黄色荧光蛋白。结果显示,转染后的柔嫩艾美耳球虫可以在体内发育阶段及卵囊阶段均表达黄色荧光蛋白,表明本研究建立了球虫体内瞬时转染系统,能简便可靠的检测黄色荧光蛋白在球虫内的瞬时表达。为进一步球虫的稳定转染提供了有力的技术支持。Genetic manipulation is a poweful tool for investigating the biology of Eimeria species. In the current study, sporozoites of Eimeria tenella were electroporated with the linearised transfection vec- tor pEtHEA,and then inoculated into the ileocaecal opening of 4-day-old chickens via the cloacal route. Various endogenous stages of E. tenella and progeny oocysts were examined using fluorescent microscope to find some fluorescent endogenous stages and oocysts of E. tenella. The Results demonstrated that an in v/vo transient expression system of E. tenella was successfully developed. YFP-expressing various stages and oocysts were obviously seen in the mucosal membranes of infected chickens and faeces, respectively. This in vivo transient transfection system is helpful for the further stable transfection of Eimeria species.

关 键 词：体内瞬时转染系统 柔嫩艾美耳球虫 黄色荧光蛋白 

分 类 号：S852.723[农业科学—基础兽医学]