斯氏副柔线虫半胱氨酸蛋白酶Pj-CPR基因的克隆及原核表达  

Cloning and Prokaryotic Expression of Pj-CPR Gene of Parabronema skrjabini

作　　者：王文龙 冯陈晨 王金玲 呼和巴特尔 刘春霞[2] 

机构地区：[1]内蒙古农业大学兽医学院/农业部动物疾病临床诊疗技术重点实验室,内蒙古呼和浩特010018 [2]内蒙古农业大学生命科学学院,内蒙古呼和浩特010018

出　　处：《动物医学进展》2018年第3期30-34,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31260603);内蒙古自然科学基金项目(2016MS0341)

摘　　要：为了筛选斯氏副柔线虫抗原基因用于血清学诊断和免疫防治研究,对斯氏副柔线虫半胱氨酸蛋白酶Pj-CPR基因序列进行了生物信息学分析,RT-PCR扩增获得了Pj-CPR基因,构建重组表达质粒pETCPR后,转入大肠埃希菌BL21(DE3)中并诱导表达获得重组蛋白rCPR。SDS-PAGE和Western blot检测结果表明,重组蛋白rCPR大小为17.6ku,主要以包涵体的形式表达,能与感染斯氏副柔线虫的骆驼血清中IgG特异性结合,具有良好的抗原活性,可用于斯氏副柔线虫病血清学诊断方法和免疫防控研究。In order to screen the antigen genes of Parabronema skrjabini for serological diagnosis and im- mune control of parabronemiasis,in this study,the eystine protease from Parabronema skrjabini （Pj-CPR） gene was analyzed by bioinformatics. RT-PCR was used to obtain Pj-CPR. The recombinant plasmid pETCPR was transformed into E.coli BL21 （DE3） and induced to express the recombinant protein rCPR. The results of SDS-PAGE and Western blot showed that the rCPR size of the recombinant protein was 17.6 ku,which was mainly expressed in the form of inclusion bodies and could specifically bind to IgG in the camel serum infected with parasites,and had good antigenic activity.The results could lay the founda- tion for the study of serological diagnosis and immune control of parabronemiasis.

关 键 词：斯氏副柔线虫 半胱氨酸蛋白酶 原核表达 骆驼 

分 类 号：S855.9[农业科学—临床兽医学]