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| H9N2亚型禽流感病毒NA基因的原核表达及其免疫反应性分析 | |
| 引用本文： | 易华山,侯顺利,独军政,胡永浩,包世俊,常惠芸.H9N2亚型禽流感病毒NA基因的原核表达及其免疫反应性分析[J].甘肃农业大学学报,2008,43(3). |
| 作者姓名： | 易华山 侯顺利 独军政 胡永浩 包世俊 常惠芸 |
| 作者单位： | 1. 中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,农业部畜禽病毒学重点开放实验室,甘肃,兰州,730046;甘肃农业大学动物医学院,甘肃,兰州,730070 2. 中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,农业部畜禽病毒学重点开放实验室,甘肃,兰州,730046;兰州军区疾病预防控制中心,甘肃,兰州,730020 3. 中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,农业部畜禽病毒学重点开放实验室,甘肃,兰州,730046 4. 甘肃农业大学动物医学院,甘肃,兰州,730070 |
| 基金项目： | 农业部畜禽病毒学重点开放实验室开放课题 |
| 摘 要： | 采用PCR方法扩增H9N2亚型禽流感病毒A/Ck/GS/2/99株NA基因(1 400 bp),克隆至pET-28a(+)载体上,构建重组原核表达载体pET-NA,转化E.coliBL21(DE3)感受态细胞,以终浓度0.8 mmol/L的IPTG诱导表达8 h.表达产物经SDS-PAGE和Western-blot分析表明:该载体能高效表达NA蛋白,相对分子质量约52KD,纯化产物能与N2亚型的AIV鸡血清发生特异性反应. |
| 关 键 词： | 禽流感 血凝素 原核表达 H9N2 免疫反应性 |
Prokaryotic expression of avian influenza virus NA Gene(H9N2)and its immunoreactivity analysis |
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| YI Hua-shan,HOU Shun-li,DU Jun-zheng,HU Yong-hao,BAO Shi-jun,CHANG Hui-yun.Prokaryotic expression of avian influenza virus NA Gene(H9N2)and its immunoreactivity analysis[J].Journal of Gansu Agricultural University,2008,43(3). | |
| Authors: | YI Hua-shan HOU Shun-li DU Jun-zheng HU Yong-hao BAO Shi-jun CHANG Hui-yun |
| Abstract: | The NA gene with a length of 1 400 bp was amplified by PCR from A/Ck/GS/2/99(H9N2)strains of avian influenza virus.The correct NA gene was inserted into the expression vector pET-28a( ) to obtain the recombinant plasmid pET-NA.The target gene was successfully expressed in the host cell E.coli BL21(DE3)induced with 0.8 mmol/L IPTG for 8 hours.The NA protein with relative molecular mass of approximately 52 000 was observed on the SDS-PAGE gel.The immunoreactivity of expressed protein was analyzed by Western blot using positive serum of avian influenza virus H9N2 subtype.The results showed that recombinant NA protein can bind to positive serum of N2 subtype. |
| Keywords: | avian influenza virus NA prokaryotic expression H9N2 immunoreactivity |
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