摘要
成功克隆了FMDV太保毒株3ABC全基因片段,并将ABC、3AB、3BC片段插入pGEX-4T-1表达载体构建了重组表达质粒,经Western blotting 分析表明,表达的3ABC、3AB蛋白与FMDV阳性血清有反应性,表达的3BC蛋白反应性差。以纯化的3ABC、3AB表达蛋白为抗原建立间接ELISA方法,对背景清楚的试验牛血清进行检测。结果3ABC、3AB表达蛋白与空白对照组(C组)、灭活疫苗反复免疫组(CI组)和部分免疫后攻毒组(I组)牛血清均不发生反应,与未免疫直接攻毒组(D组)和部分I组血清均发生反应;未免疫直接攻毒组3ABC、3AB表达蛋白抗体持续时间几乎相同,至少在90d以上;免疫后攻毒组3ABC比3AB表达蛋白抗体持续时间长,说明3ABC表达蛋白更适合用于FMDV感染动物的检疫。
A gene fragment of 3ABC gene of the foot-and-mouth disease virus (FMDV) TaiBao strain was successfullycloned, and 3ABC, 3AB, 3BC expressing plasmids were constructed by inserting the target gene fragments into pGEX-4T-1 vectors respectively. The expression proteins were analysed by Western blot methods. The result showed that theexpressed 3ABC and 3AB proteins strongly reacted with FMDV positive serum while the reaction of the 3BC protein withthe FMDV positive serum was poor. An indirect ELISA was set up using purified 3ABC and 3AB proteins as antigen andtested against 4 groups of cattle serum: control group, vaccinated-group, infected-group and infected-group after vaccination.Negative results were obtained using the control group, the vaccinated group and a part of the infected-group aftervaccination. On the contrary, positive results were obtained in the infected group and a part of the infected-group aftervaccination. In the infected group, the duration of the antibody against 3ABC and 3AB were the same and have lasted forat least 90 days. The duration of the antibody against 3ABC is longer than that against 3AB in the infected-group aftervaccination. This indicated that the expression 3ABC protein is more suitable for using as an antigen to differentiate theconvalescent and vaccinated animals.
出处
《中国农业科学》
CAS
CSCD
北大核心
2004年第7期1065-1070,共6页
Scientia Agricultura Sinica
基金
科技部社会公益研究专项基金资助项目(2003DIA7J033)
