摘要
利用限制性酶切从重组质粒pShuttle-CMV-VP中得到猪O型口蹄疫病毒VP1(21-60)-(141-160)-(200-213)位氨基酸的基因。将此多抗原表位基因克隆至原核高效表达载体pET43.1 a(+),在E.coliBL21中用IPTG诱导表达了含有猪口蹄疫病毒多抗原表位的融合蛋白,并用镍柱亲和层析法获得了纯化蛋白。W estern-b lot结果表明融合蛋白可被猪O型口蹄疫病毒标准阳性血清所识别,从而为进一步研究FMDV多表位抗原的免疫特性和诊断方法奠定了基础。
Amino acids （21-60） - （141-160） - （200-213） coding region of swine foot-and-mouth disease virus （FMDV） serotype O VPI was successfully obtained from recombinant plasmid pShuttle-CMV-VP by restriction enzymes and cloned into prokaryotic expression vector pET43.1a （ ＋ ）. The fusion protein was highly expressed in E. coli BL21 induced by IPTG. It could be purified efficiently with Ni^＋ affinity chromatography column. Western-blot analysis showed that the fusion protein was able to react with swine polyclonal antibody against swine foot-and-mouth disease virus serotype O. It provided fundamental data and materials for the further investigation on immunogenicity of multiple epitopes vaccine of FMDV.
出处
《畜牧与兽医》
北大核心
2006年第10期1-3,共3页
Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金(B0270990)
新世纪优秀人才支持计划(NCET-04-0502)
关键词
口蹄疫病毒
表位
原核表达
foot-and-mouth disease virus
epitope
prokaryotic expression
