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甲型H1N1流感病毒NA、PB2和PA表位抗原区段的克隆和表达
引用本文:宋晓国,王国华,朱翠侠,戴振华,修冰水,何竞,陈坤,张向颖,凌世淦,杨静,王升启,张贺秋,冯晓燕. 甲型H1N1流感病毒NA、PB2和PA表位抗原区段的克隆和表达[J]. 生物技术通讯, 2010, 21(1): 17-21. DOI: 10.3969/j.issn.1009-0002.2010.01.004
作者姓名:宋晓国  王国华  朱翠侠  戴振华  修冰水  何竞  陈坤  张向颖  凌世淦  杨静  王升启  张贺秋  冯晓燕
作者单位:1. 军事医学科学院,基础医学研究所,北京,100850
2. 军事医学科学院,放射和辐射医学研究所,北京,100850
摘    要:目的:分析比对甲型H1N1流感病毒神经氨酸酶(NA)、聚合酶B2(PB2)和聚合酶A(PA)抗原的序列,克隆表达保守的和变异的表位抗原区段,为免疫学诊断试剂的研究提供候选抗原。方法:采用BioSun生物学软件比较新近公布的A/H1N1流感病毒和我国猪流感病毒浙江株NA、PB2和PA抗原序列,并预测筛选NA、PB2和PA抗原表位保守区段与变异区段,采用PCR逐步合成法合成NA、PB2和PA表位抗原区段基因序列,并利用原核表达载体pBVIL1进行克隆表达。结果:筛选的保守表位抗原区段和变异表位抗原区段为NA/135~180aa、NA/310~350aa、PB2/1~80aa、PA/41~90aa、PA/231~280aa和PA/341~400aa;获得6条表位抗原区段基因序列,且6条表位抗原区段均获得了高效表达,得到纯化后的抗原。结论:获得了A/H1N1流感病毒NA、PB2和PA表位抗原区段,为进一步研制特异的甲型流感病毒快速诊断试剂提供了抗原储备。

关 键 词:甲型H1N1流感病毒  神经氨酸酶  聚合酶B2  聚合酶A  表位抗原区段  克隆  表达

Cloning and Expression of NA,PB2 and PA of Novel H1N1 Influenza A Virus
SONG Xiao-Guo,WANG Guo-Hua,ZHU Cui-Xia,DAI Zhen-Hua,XIU Bing-Shui,HE Jing,CHEN Kun,ZHANG Xiang-Ying,LING Shi-Gan,YANG Jing,WANG Sheng-Qi,ZHANG He-Qiu,FENG Xiao-Yan. Cloning and Expression of NA,PB2 and PA of Novel H1N1 Influenza A Virus[J]. Letters in Biotechnology, 2010, 21(1): 17-21. DOI: 10.3969/j.issn.1009-0002.2010.01.004
Authors:SONG Xiao-Guo  WANG Guo-Hua  ZHU Cui-Xia  DAI Zhen-Hua  XIU Bing-Shui  HE Jing  CHEN Kun  ZHANG Xiang-Ying  LING Shi-Gan  YANG Jing  WANG Sheng-Qi  ZHANG He-Qiu  FENG Xiao-Yan
Affiliation:SONG Xiao-Guoa,WANG Guo-Huaa,ZHU Cui-Xiaa,DAI Zhen-Huaa,XIU Bing-Shuia,HE Jinga,CHEN Kuna,ZHANG Xiang-Yinga,LING Shi-Gana,YANG Jingb,WANG Sheng-Qib,ZHANG He- Qiua,FENG Xiao-Yana a.Institute of Basic Medical Sciences,b.Institute of Radiation Medicine,Academy of Military Medical Sciences,Beijing 100850,China
Abstract:Objective:In order to provide the candidate antigens for immunological diagnosis,to analysis the sequence of neuraminidase(NA),polymerase B2(PB2)and polymerase A(PA)of H1N1 influenza A virus,and to clone and express the conservative and variable antigen epitope segments of the NA,PB2 and PA.Methods:The amino acid sequences of the newly published NA,PB2 and PA,and Zhejiang swine influenza virus NA,PB2 and PA were analyzed and aligned using the BioSun software.The genes of antigen epitope segments were synthe...
Keywords:H1N1 influenza A virus  neuraminidase  polymerase B2  polymerase A  antigen epitope segment  cloning  expression  
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