首页 | 本学科首页   官方微博 | 高级检索  
     

禽流感 HA 基因密码子优化的 DNA 疫苗构建及其免疫原性的初步研究
引用本文:张春华,张璐,王世霞,李军,黄祖瑚,卢山. 禽流感 HA 基因密码子优化的 DNA 疫苗构建及其免疫原性的初步研究[J]. 中国医药生物技术, 2007, 2(2): 110-113
作者姓名:张春华  张璐  王世霞  李军  黄祖瑚  卢山
作者单位:1. 210029,南京,江苏省人民医院中美疫苗研究中心
2. 美国马萨诸塞州大学医学院核酸疫苗研究室;210029,南京,江苏省人民医院中美疫苗研究中心
摘    要: 目的 构建H5 N1亚型禽流感病毒株密码子优化的血凝素(HA)DNA疫苗并研究其免疫原性。方法 应用MacVector软件分析H5N1亚型禽流感病毒A/Viet Nam 1203/04株HA(H5-VN HA)基因序列,对这些序列进行密码子优化处理,人工合成优化后的H5-VN HA基因。将密码子优化H5-VN HA基因与pSW3891质粒连接,并以人组织纤维蛋白溶酶原激活剂(tPA)信号肽取代H5-VN HA信号肽,构建成表达H5-VN HA基因的重组质粒,命名为HA-VN.tPA(即密码子优化的H5-VN HA基因DNA疫苗)。以HA-VN.tPA转染293T细胞,应用蛋白质印迹法检测转染细胞中HA蛋白的表达。以HA-VN.tPA对2只新西兰兔进行DNA免疫,用ELISA方法检测分析免疫后兔血清中HA特异性抗体的产生。结果 密码子优化的H5-VN HA基因中哺乳动物细胞偏爱的密码子频率高于野生型H5-VN HA基因。蛋白质印迹法分析结果表明,HA-VN.tPA转染293T细胞后,细胞裂解液中检测出HA蛋白的表达。ELISA分析结果表明,以HA-VN.tPA免疫3次后,实验兔体内产生了较高水平的HA特异性抗体(血清中平台期抗体滴度达1:13 500)。结论 成功构建了H5N1亚型禽流感病毒密码子优化的HA 基因DNA疫苗,该疫苗可引导免疫后宿主体内产生高滴度特异性抗体。

关 键 词:疫苗,DNA  流感病毒,禽  血凝素,病毒  密码子
收稿时间:2007-02-07
修稿时间:2007-02-07

Immunogenicity of a codon optimized DNA vaccine expressing avian flu HA antigen
ZHANG Chun-hua,ZHANG Lu,WANG Shi-xia,LI Jun,HUANG Zuhu,LU Shan. Immunogenicity of a codon optimized DNA vaccine expressing avian flu HA antigen[J]. Chinese Medicinal Biotechnology, 2007, 2(2): 110-113
Authors:ZHANG Chun-hua  ZHANG Lu  WANG Shi-xia  LI Jun  HUANG Zuhu  LU Shan
Abstract:Objective To study the immunogenicity of a DNA vaccine expressing the codon optimized HA gene from an H5N1 avian flu viral isolate, A/Viet Nam1203/04 (H5-VN). Methods The codon usage of the wild type H5-VN HA gene was analyzed with the MacVector software against the codon preference for Homo sapiens. Synthetic codon optimized H5-VN A HA gene was cloned into DNA vaccine vector pSW3891, and H5-VN HA gene leader sequence was substituted with a human tissue plasminogen activator (tPA) leader sequence. The pSW3891 vector with the tPA leader, which was named HA-VN.tPA, was transfected into 293T cells. And the expression of HA gene in the transfected cells was detected by Western blot. Two New Zealand rabbits were immunized with electroporation method, then the anti-HA specific antibodies in the serum of the rabbits were evaluated by ELISA. Results In the codon optimized H5-VN HA gene, the frequency of the mammal cell preferred codons was higher than that of the wild type H5-VN HA gene. By Western blotting, the expression of HA protein was detected in 293T cell lysate transfected by HA-VN.tPA. ELISA showed that anti-H5 specific antibody was produced after three-time vaccinations Conclusion A H5-VN HA DNA vaccine was successfully constructed with proven immunogenicity in eliciting high-titre avian flu HA specific antibody response.
Keywords:Vaccines   DNA  Influenza   avian  Hemagglutinins   viral  Codon
本文献已被 CNKI 维普 万方数据 等数据库收录!
点击此处可从《中国医药生物技术》浏览原始摘要信息
点击此处可从《中国医药生物技术》下载免费的PDF全文
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号