摘要
从感染驴白细胞的马传贫驴白细胞弱毒疫苗株前病毒DNA中克隆了编码基质蛋白 (p1 5 )的基因 ,并在大肠杆菌中进行了表达。所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。在间接ELISA和免疫印迹试验中 ,重组的基质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。这表明该重组蛋白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中。
Gene coding for matrix protein (p15) of donkey leukocyte 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. Protein was expressed in a soluble form as fusion protein.It was purified by immobilized metal ion affinity chromatography under native conditions because there was a 6 histidines tag at its amino acid terminus.Recombinant matrix protein showed reactivity to EIAV positive serum samples and no reactivity to normal horse sera in indirect ELISA and immunoblot assay.This demonstrates that recombinant p15 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 donkey leukocyted attenuated vaccine strain of equine infectious anemia virus.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2003年第1期1-4,共4页
Chinese Journal of Preventive Veterinary Medicine
关键词
基质蛋白
马
传染性贫血病
马传贫驴白细胞弱毒疫苗株
基因克隆
基因表达
P15基因
Equine infectious anemia
Recombinant p15 matrix protein
Donkey leukocy teadapted attenuated vaccine strain of equine infectious anemia virus
