| H5N1型禽流感模型侵染病毒的构建与鉴定 |
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| 引用本文： | 万佳,胡建和,王青,杭柏林,王三虎,刘兴友.H5N1型禽流感模型侵染病毒的构建与鉴定[J].西北农林科技大学学报(社会科学版),2010(3):23-27. |
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| 作者姓名： | 万佳 胡建和 王青 杭柏林 王三虎 刘兴友 |
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| 作者单位： | 河南科技学院动物科学学院;郑州大学临床学院; |
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| 基金项目： | 河南省高校杰出科研人才创新工程项目(2006KYCX020) |
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| 摘 要： | 【目的】构建含有H5N1型禽流感病毒HA、NA的侵染模型病毒,并对构建的模型病毒进行侵染活性及特异性检测。【方法】采用PCR方法扩增出H5N1型AIVHA和NA基因,经纯化回收后将其分别克隆至真核表达载体pcDNA4.0中,构建pcDNA-HA、pcDNA-NA。然后采用磷酸钙共转染方法,将pcDNA-HA、pcDNA-NA和含有LUC报告系统的HIV骨架表达载体pNL4-3.Luc.R-E-,共同转染293T细胞,构建HIV/HA-NA模型病毒,对HIV/HA-NA模型病毒进行细胞侵染活性及特异性检测。【结果】HA、NA基因片段真核表达载体pcDNA-HA、pcDNA-NA与HIV骨架载体在293T细胞中成功表达组装,获得了HIV/HA-NA侵染模型病毒;敏感细胞特异性检测及侵染活性分析表明,HIV/HA-NA模型病毒与其野毒具有共同的敏感细胞。报告系统数据显示,模型病毒可以定量检测其对细胞的侵染程度。【结论】成功构建HIV/HA-NA模型侵染病毒,可以模拟H5N1病毒的侵染过程,定量报告病毒的侵染程度,并且该模型病毒不具有增殖能力,安全可靠,可以方便地用于普通实验室AIV抗体检测和特定药物的筛选。
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| 关 键 词： | 禽流感 模型病毒 磷酸钙共转染 报告基因 |
Construction and identification of infectious model virus of AIV strain H5N1 |
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| WAN Jia,HU Jian-he,WANG Qing,HANG Bo-lin,WANG San-hu,LIU Xing-you.Construction and identification of infectious model virus of AIV strain H5N1[J].Journal of Northwest Sci-Tech Univ of Agr and,2010(3):23-27. |
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| Authors: | WAN Jia HU Jian-he WANG Qing HANG Bo-lin WANG San-hu LIU Xing-you |
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| Institution: | WAN Jia1,2,HU Jian-he1,WANG Qing1,HANG Bo-lin1,WANG San-hu1,LIU Xing-you1(1 School of Animal Science , Veterinary Medicine,Henan Institute of Science , Technology,Xinxiang,Henan 453003,China,2 Clinical Medical College,Zhengzhou University,Zhengzhou,Henan 450052,China) |
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| Abstract: | Objective] HA and NA of Avian influenza virus strain H5N1 were used to construct a model virus of AIV.The activity and specificity of the model virus were tested respectively.Method] HA and NA genes of AIV H5N1 were amplified and subcloned into pcDNA4.0.The recombinant plasmids were named pcDNA-HA and pcDNA-NA respectively.Using calcium phosphate cotransfection method,these plasmids together with HIV backbone pNL4-3.Luc.R-E-were transfected into 293T cell lines.The model virus was named HIV/HA-NA.The acti... |
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| Keywords: | Avian influenza virus model virus calcium phosphate cotransfection method gene reported |
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