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| 肠道病毒71型SHZH03株VP1基因在毕赤酵母中的表达 | |
| 引用本文： | 周世力,李琳琳,何雅青.肠道病毒71型SHZH03株VP1基因在毕赤酵母中的表达[J].华中师范大学学报(自然科学版),2007,41(2):259-262. |
| 作者姓名： | 周世力 李琳琳 何雅青 |
| 作者单位： | 1. 江汉大学,医学与生命科学学院,武汉,430056 2. 中国疾病预防与控制中心病,毒基因工程国家重点实验室,北京,100052 3. 深圳市疾病预防与控制中心,广东,深圳,518001 |
| 摘 要： | 利用逆转录聚合酶链式反应(RT-PCR)方法扩增得到肠道病毒71型(EV71SHZH03)外壳蛋白VP1基因,经序列测定证实后,构建重组表达质粒VP1/pPIC9K,转化Pichia pastoris 酵母宿主菌Gs115,以Myc-Tag多克隆抗体作为一抗,利用双层滤膜法筛选酵母转化子.甲醇诱导表达.SDS-PAGE分析显示:表达产物的分子量约为30000,与天然VP1大小一致,ELISA实验表明,表达上清液可与EV71患者急性感染期血清呈阳性反应,表明重组蛋白VP1具有免疫原性. |
| 关 键 词： | 肠道病毒71型SHZH03株 VP1 毕赤酵母 肠道 病毒 基因 毕赤酵母 诱导表达 Pichia Pastoris 免疫原性 重组蛋白 阳性反应 血清 急性感染期 患者 上清液 实验 ELISA 大小 分子量 显示 分析 甲醇 |
| 文章编号： | 1000-1190（2007）02-0259-04 |
| 修稿时间： | 2006-12-19 |
Expression of VP1 of Enterovirus71 SHZH03 in Pichia Pastoris |
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| ZHOU Shili,LI Linlin,HE Yaqin.Expression of VP1 of Enterovirus71 SHZH03 in Pichia Pastoris[J].Journal of Central China Normal University(Natural Sciences),2007,41(2):259-262. | |
| Authors: | ZHOU Shili LI Linlin HE Yaqin |
| Institution: | 1. School of Medical and Life science, Jianghan University, Wuhan 430056 2. The State Key Laboratory for Molecular Virology and Gene Engineering, Chinese Center for Disease Control and Prevention, Beijing 10052; 3. Center for Disease Control and Prevention of Shenzhen, Shenzhen, Gangdong 518001 |
| Abstract: | The gene encoding the entire capsid protein VP1 of Enterovirus 71SHZH03 was amplified by RT-PCR. After confirmed by sequence analysis, VP1 was expressed in Pichia pastoris (GS115). In situ two film method was used to screen Pichia pastoris transformants expressing high level protein of VP1. SDS-PAGE showed that the gene vp1 could express a 30000 equivalent to the natural protein. ELISA analysis of the culture supernatants confirmed that the protein had the desired immunogenicity of EV71. |
| Keywords: | Enterovirus 71SHZH03 VP1 Pichia Pastoris |
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