摘要
从感染驴白细胞的马传染性贫血弱毒疫苗株前病毒DNA中克隆了编码核衣壳蛋白 (pll)的基因 ,在大肠杆菌中得到了表达 ,而表达的蛋白是一种可溶性的融合蛋白 ,其氨基端带有 6个组氨酸的标签 ,因此可以用固定化金属离子亲和层析法在非变性条件下进行纯化。经间接ELISA和免疫印迹试验检测 ,这种表达的融合蛋白可与马传贫阳性血清样品发生反应 ,而与健康马血清无任何反应 ,显示其具有良好的抗原性和特异性 ,可用于马传贫弱毒疫苗株在体内外复制及在接种马体内免疫应答的研究。
The gene coding for the nucleogapsid protein (pll) of the attenuated vaccine strain of equine infectious anemia virus (EIAV) was cloned from its proviral DNA obtained from the vaccine strain infected donkey leukocyte cultures, and expressed in E.coli. The protein was expressed in a soluble form and as a fusion protein. There was a 6 histidines tag at its amino acid terminus. It was purified by immobilized metal ion affinity chromatography under native conditions. Recombinant nucleoprotein protein showed reactivity to EIAV positive serum samples and no reactivity to normal horse sera in indirect ELISA and immuoblot assay. This demonstrates that recombinant pll nucleoprotein protein has very good antigenicity and specificity and may be used for research on the in vitro and in vivo replication and immune responses in the vaccinated horses of the attenuated vaccine strain of equine infectous anemia virus.;
出处
《中国生物工程杂志》
CAS
CSCD
2004年第4期67-70,共4页
China Biotechnology
关键词
马传染性贫血病毒
弱毒疫苗株
克隆
DNA
核衣壳蛋白
基因
表达
Attenuated vaccine strain of equine infectious anemia virus Recombinant nucleoprotein protein pll Expression
