夹心ELISA法测定基因重组球虫表达异源蛋白的水平  

The Heterologous Protein Expression Level in Recombinant Eimeria was Detected by Sandwich ELISA

作　　者：罗伏兵 徐发荣 王慧强 马志军 汤新明[2] 刘贤勇 索勋[2] 刘晓冬 LUO Fu-bing;XU Fa-rong;WANG Hui-qiang;MA Zhi-jun;TANG Xin-ming;LIU Xian-yong;SUO Xun;LIU Xiao-dong(Beijing Center for Animal Disease Control and Prevention,Beijing 102629,China;College of Veterinary Medicine,China Agricultural University,Beijing 100193,China)

机构地区：[1]北京市动物疫病预防控制中心,北京大兴102629 [2]中国农业大学动物医学院,北京海淀100193

出　　处：《中国兽医杂志》2020年第8期36-38,I0005,共4页Chinese Journal of Veterinary Medicine

基　　金：北京市家禽产业创新团队(BAIC04-2019)。

摘　　要：为了评估基因重组球虫表达异源蛋白的水平,本文以表达黄色荧光蛋白的基因重组球虫为模型,建立检测基因重组球虫可溶性蛋白中黄色荧光蛋白含量的夹心ELISA方法。基于此,我们检测了不同启动子调控蛋白表达的基因重组球虫中黄色荧光蛋白的含量,发现His4启动子调控表达的黄色荧光蛋白占球虫可溶性蛋白的3.1‰,而SAG13启动子调控表达的黄色荧光蛋白占球虫可溶性蛋白的9.4‰。结果表明,夹心ELISA法可用于检测基因重组球虫中异源蛋白的表达水平,并具有较高的灵敏性。本方法为评估基因重组球虫表达异源抗原的含量和优化基因重组球虫作为疫苗载体的研究提供了科学依据,同时也为其他类型疫苗载体或相关研究提供借鉴。In order to evaluate the expression level of heterologous protein in recombinant Eimeria,a sandwich ELISA was developed based on detection of the expression level of enhanced yellow fluorescent protein(EYFP)in the soluble protein of recombinant Eimeria.We compared the expression level of EYFP,which was regulated by histone 4(His4)and surface antigen 13(SAG13)promoter.We found that EYFP regulated by His4 promoter accounted for 3.1‰,whereas,EYFP regulated by SAG13 promoter accounted for 9.4‰of the recombinant Eimeria soluble proteins.These results demonstrated that sandwich ELISA is a sensitive method which could be used to detect the specific protein in a mixture and lay the foundation for evaluating the expression level of heterologous antigen in recombinant Eimeria and optimizing the recombinant Eimeria as a vaccine vector.The results also have implications on optimizing other vaccine vectors or related research.

关 键 词：基因重组球虫 黄色荧光蛋白 ELISA 表达水平 疫苗载体 

分 类 号：R382.3[医药卫生—医学寄生虫学]