| 非洲猪瘟病毒VP73蛋白主要抗原表位区的原核表达及多克隆抗体的制备 |
| |
| 引用本文： | 李洪利,王君玮,张维,吴晓东,赵永刚,李慧,李娟,包静月,曹金山,王志亮.非洲猪瘟病毒VP73蛋白主要抗原表位区的原核表达及多克隆抗体的制备[J].中国畜牧兽医,2012,39(10):7-10. |
| |
| 作者姓名： | 李洪利 王君玮 张维 吴晓东 赵永刚 李慧 李娟 包静月 曹金山 王志亮 |
| |
| 作者单位： | 1. 中国动物卫生与流行病学中心,山东青岛 266114;2. 内蒙古农业大学兽医学院,内蒙古呼和浩特 010018;3. 山东农业大学兽医学院,山东泰安 271000 |
| |
| 基金项目： | 国家公益性行业(农业)科研专项经费 |
| |
| 摘 要： | 构建重组原核表达载体pET32a-VP73,将pET32a-VP73转化BL21感受态细胞,经IPTG诱导,VP73蛋白主要抗原表位区可稳定高效的表达,SDS-PAGE结果表明,IPTG终浓度为1.0 mmol/L,诱导5 h蛋白质表达量最高,表达蛋白为融合蛋白,分子质量约为42 ku。蛋白质纯化后,经SDS-PAGE及Western blotting鉴定,确定表达产物为非洲猪瘟病毒VP73主要抗原表位区融合蛋白。将纯化的蛋白质免疫新西兰大白兔,免疫前后收集血清。用间接ELISA方法测定血清抗体效价,并以非洲猪瘟阳性血清、兔免疫前后血清及猪瘟、猪繁殖与呼吸综合征、猪伪狂犬病阳性血清为一抗,确定该蛋白质的特异性。结果表明,制备的抗血清效价达到1∶1024000,能与纯化的VP73主要抗原表位区蛋白质发生反应。制备的多克隆抗体为VP73蛋白的免疫学研究和非洲猪瘟血清学诊断奠定了基础。
|
| 关 键 词： | 非洲猪瘟 VP73 原核表达 多克隆抗体 |
| 收稿时间： | 2012-02-22 |
Prokaryotic Expression of Major Antigenic Epitope Region of African Swine Fever Virus VP73 Protein and Preparation of Polyclonal Antibody |
| |
| LI Hong-li
,
WANG Jun-wei
,
ZHANG Wei
,
WU Xiao-dong
,
ZHAO Yong-gang
,
LI Hui
,
LI Juan
,
BAO Jing-yue
,
CAO Jin-shan
,
WANG Zhi-liang.Prokaryotic Expression of Major Antigenic Epitope Region of African Swine Fever Virus VP73 Protein and Preparation of Polyclonal Antibody[J].China Animal Husbandry & Veterinary Medicine,2012,39(10):7-10. |
| |
| Authors: | LI Hong-li WANG Jun-wei ZHANG Wei WU Xiao-dong ZHAO Yong-gang LI Hui LI Juan BAO Jing-yue CAO Jin-shan WANG Zhi-liang |
| |
| Institution: | 1. China Animal Health and Epidemiology Center, Qingdao 266114, China;2. College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China;3. College of Veterinary Medicine, Shandong Agricultural University, Tai’an 271000, China |
| |
| Abstract: | Constructed recombinant prokaryotic expression vector pET32a-VP73 was transformed into E.coli BL21, major antigenic epitope region of VP73 protein was induced stably and efficiently by IPTG, the result of SDS-PAGE showed, when final concentration of IPTG was 1.0 mmol/L, and pET32a-VP73 was induced 5 h, expression of major antigenic epitope region of VP73 protein was highest, the expressed protein was fusion protein, molecular weight was approximately 42 ku. The protein was identified as major antigenic epitope region of VP73 protein by SDS-PAGE and Western blotting. The purified major antigenic epitope region of VP73 protein was injected to New Zealand rabbits, serum was collected before and after injection. Antibody titer was determined by ELISA. Specificity of this protein was determined by ELISA whose primary antibody were ASF positive serum, rabbits serum before and after injection and positive serum of CSF, PRRS, pig pseudorabies. The result showed the titer of antiserum reached up to 1∶1024000, and gave rise to reactions with VP73 major antigenic region protein. The polyclonal antibody built the foundation for immunology research of VP73 and serological diagnosis of African swine fever. |
| |
| Keywords: | African swine fever VP73 prokaryotic expression polyclonal antibody |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《中国畜牧兽医》浏览原始摘要信息 |
| 点击此处可从《中国畜牧兽医》下载免费的PDF全文 |
|
