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| 流感病毒RNA聚合酶串联亲和纯化方法的建立 | |
| 引用本文： | 李印,邓国华,崔佳莹,于洋,丁睛薇,陈化兰.流感病毒RNA聚合酶串联亲和纯化方法的建立[J].中国预防兽医学报,2011,33(11). |
| 作者姓名： | 李印 邓国华 崔佳莹 于洋 丁睛薇 陈化兰 |
| 作者单位： | 1. 中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室 兽医生物技术国家重点实验室,黑龙江哈尔滨,150001 2. 华南农业大学,广东广州,510642 3. 湖南农业大学,湖南长沙,410127 |
| 基金项目： | 国家十五重点科技攻关课题 |
| 摘 要： | 为建立流感病毒RNA聚合酶的串联亲和纯化方法,本实验由鸡胚尿囊液中提取病毒核酸,经过RT-PCR和常规PCR方法扩增禽流感病毒株A/Duck/Guangxi/35/01 (H5N1)的PB1、PB2和PA的cDNA.分别将PB2基因连接于pcDNATAP载体中,PB1和PA基因连接于pcDNA3A载体中构建了pcDNA-35PB2TAP、pcDNA-35PB1和pcDNA-35PA重组质粒.将构建的真核表达重组质粒共转染293T细胞,经过串联亲和纯化方法获得流感病毒依赖RNA的RNA聚合酶复合体(RdRp).利用RNA体外合成试验验证获得的RdRp具有生物学活性. |
| 关 键 词： | H5亚型禽流感病毒 RNA聚合酶复合体 串联亲和纯化 |
Tandem affinity purification of influenza virus RNA polymerase |
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| LI Yin,DENG Guo-hua,CUI Jia-ying,YU Yang,DING Qing-wei,CHEN Hua-lan.Tandem affinity purification of influenza virus RNA polymerase[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(11). | |
| Authors: | LI Yin DENG Guo-hua CUI Jia-ying YU Yang DING Qing-wei CHEN Hua-lan |
| Institution: | LI Yin1,DENG Guo-hua1,CUI Jia-ying1,YU Yang2,DING Qing-wei3,CHEN Hua-lan1(1.State Key Laboratory of Veterinary Biotechnology,Animal Influenza Laboratory of the Ministry of Agricultural,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China,2.South China Agricultural University,Guangzhou 510642,3.Hunan Agricultural University,Changsha 410127,China) |
| Abstract: | To express and purify the influenza virus RNA polymerase,the total RNA of H5 subtype avian influenza strain A/Duck/Guangxi/35/01(H5N1) was extracted from allantoic fluid and the genes of PB1,PB2 and PA were amplified by RT-PCR.The recombinant plasmids of pcDNA-35PB2TAP,pcDNA-35PB1 and pcDNA-35PA were constructed based an eurokaryotic expression vetors of pcDNATAP and pcDNA3A,respectively.Furthermore,the RNA-dependent RNA polymerase complex(RdRp) of the virus expressed in the co-transfected 293T with those r... |
| Keywords: | H5 subtype avian influenza virus RNA polymerase tandem affinity purification |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |