|
|||||
|
|
| 人细小病毒B19病毒样颗粒的制备 | |
| 引用本文： | 邹小辉,董流昕,宋敬东,屈建国,于修平,鲁茁壮,洪涛. 人细小病毒B19病毒样颗粒的制备[J]. 生物工程学报, 2009, 25(4): 575-579 |
| 作者姓名： | 邹小辉 董流昕 宋敬东 屈建国 于修平 鲁茁壮 洪涛 |
| 作者单位： | 1. 山东大学医学院微生物学教研室,济南,250012;中国疾病预防控制中心病毒病预防控制所,北京,100052 2. 中国疾病预防控制中心病毒病预防控制所,北京,100052 3. 山东大学医学院微生物学教研室,济南,250012 |
| 基金项目： | 国家科技支撑计划(No.2008BAI56B01)资助~~ |
| 摘 要： | 采用昆虫杆状病毒表达系统,制备人细小病毒B19病毒样颗粒(VLPs)。先通过PCR方法合成细小病毒B19衣壳蛋白基因VP2,将其克隆到pFastBac1质粒,然后转化含杆状病毒穿梭载体Bacmid的E.coliDH10Bac感受态细胞,获得重组杆状病毒表达质粒Bacmid-VP2。在脂质体介导下转染Sf9昆虫细胞,包装重组杆状病毒rBac-VP2。利用rBac-VP2感染Sf9细胞表达B19VP2蛋白,通过间接免疫荧光、Western blotting等方法鉴定目的蛋白表达。采用两次超速离心的方法对表达产物进行纯化,纯化产物在透射电镜下可见直径约22nm的VLPs。本研究成功制备了人细小病毒B19的VLPs,为B19感染血清学检测方法的建立提供了参考。 |
| 关 键 词： | 细小病毒B19 VP2 杆状病毒 病毒样颗粒 |
| 收稿时间： | 2008-12-01 |
Preparation of human parvovirus B19 virus-like particles |
|
| Xiaohui Zou,Liuxin Dong,Jingdong Song,Jianguo Qu,Xiuping Yu,Zhuozhuang Lu and Tao Hong. Preparation of human parvovirus B19 virus-like particles[J]. Chinese journal of biotechnology, 2009, 25(4): 575-579 | |
| Authors: | Xiaohui Zou Liuxin Dong Jingdong Song Jianguo Qu Xiuping Yu Zhuozhuang Lu Tao Hong |
| Affiliation: | Department of Microbiology, School of Medicine, Shandong University, Jinan 250012, China; Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China;Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China;Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China;Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China;Department of Microbiology, School of Medicine, Shandong University, Jinan 250012, China;Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China;Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China |
| Abstract: | The baculovirus expression system was employed to prepare the virus-like particles (VLPs) of human parvovirus B19. The synthesized VP2 gene of B19 was inserted into the multi-cloning site (MCS) of pFastBac1 vector; the resulting plasmid was transferred to the Escherichia coli DH10Bac competent cells,which contain a baculovirus shuttle vector (Bacmid),to generate Bacmid-VP2 by site-specific transposition. Recombinant baculovirus carrying VP2 gene (rBac-VP2) was then rescued from Bacmid-VP2-transfected Sf9 ce... |
| Keywords: | parvovirus B19 VP2 baculovirus virus-like particles |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |
| 点击此处可从《生物工程学报》浏览原始摘要信息 | |
| 点击此处可从《生物工程学报》下载全文 | |