| 密码子优化对鸭坦布苏病毒E基因DNA疫苗免疫应答的增强作用 |
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| 引用本文： | 徐大伟,李国新,李雪松,李云章,李泽君.密码子优化对鸭坦布苏病毒E基因DNA疫苗免疫应答的增强作用[J].中国兽医科学,2012(5):478-482. |
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| 作者姓名： | 徐大伟 李国新 李雪松 李云章 李泽君 |
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| 作者单位： | 内蒙古农业大学兽医学院;中国农业科学院 上海兽医研究所 农业部动物寄生虫学重点开放实验室;国家家禽工程技术研究中心 |
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| 基金项目： | 公益性行业(农业)科研专项经费项目(201003012);国家自然科学基金资助项目(31172332) |
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| 摘 要： | 为提高DNA疫苗用于预防鸭坦布苏病毒病的免疫原性,将鸭坦布苏病毒的E基因密码子改造为鸭偏嗜的密码子,并将其作为免疫原基因克隆到pCAGGS载体中,构建真核表达质粒pCAGGS-OptiE。将pCAGGS-OptiE和先前构建的编码野生型E基因的表达质粒pCAGGS-E分别转染293T细胞,用间接免疫荧光和Western-blot检测E蛋白的表达情况。结果显示,密码子优化的重组质粒的蛋白表达量明显高于野生型质粒。将重组质粒pCAGGS-OptiE和pCAGGS-E用脂质体包裹后分别尾静脉注射免疫BALB/c小鼠,采用间接ELISA检测抗体的产生情况。结果显示,质粒pCAGGS-OptiE诱导的抗体水平明显高于pCAGGS-E,表明密码子优化的E基因能够提高蛋白的表达水平,进而增强机体免疫应答反应。
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| 关 键 词： | 鸭坦布苏病毒 DNA疫苗 E基因 密码子优化 |
Optimization of codon usage of E gene enhanced immune responses of DNA vaccine against duck Tembusu virus |
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| XU Da-wei,LI Guo-xin,LI Xue-song,LI Yun-zhang,LI Ze-jun.Optimization of codon usage of E gene enhanced immune responses of DNA vaccine against duck Tembusu virus[J].Veterinary Science in China,2012(5):478-482. |
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| Authors: | XU Da-wei LI Guo-xin LI Xue-song LI Yun-zhang LI Ze-jun |
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| Institution: | 2,3(1.College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China;2.Key Laboratory of Animal Parasitology of the Ministry of Agriculture/Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Shanghai 200241,China;3.National Poultry Engineering and Technology Research Center,Shanghai 200241,China) |
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| Abstract: | To improve the immunogenicity of DNA vaccines,E gene of duck Tembusu virus(DTMUV)was optimized according to the codon bias of duck.The optimized E gene(OptiE) and the wild-type E gene were cloned into the pCAGGS vector to generate the pCAGGS-OptiE and pCAGGS-E vectors,respectively.The E protein transiently expressed in the 293T cells was detected by indirect immunofluorescence assay and Western-blot.Results showed that the expression of optiE protein in vitro was increased in comparison to that of the wild-type.The immunogenicity of two constructs wrapped by liposomes was then evaluated in BALB/c mice with the antibodies detected by indirect ELISA.Results showed that the antibody level induced by pCAGGS-OptiE was higher than that did by pCAGGS-E.These results indicated that the codon optimization of E gene enhanced the immune efficacy of the DNA vaccine against DTMUV. |
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| Keywords: | duck Tembusu virus DNA vaccine E gene codon optimization |
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