| 猪繁殖与呼吸综合征病毒GP5和M基因重组腺病毒的构建及表达 |
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| 引用本文： | 高博,马玲,张念,毛燕,陈凤莲,张守峰,刘烨,张菲,扈荣良,吴健敏. 猪繁殖与呼吸综合征病毒GP5和M基因重组腺病毒的构建及表达[J]. 动物医学进展, 2012, 0(1): 23-27 |
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| 作者姓名： | 高博 马玲 张念 毛燕 陈凤莲 张守峰 刘烨 张菲 扈荣良 吴健敏 |
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| 作者单位： | 广西兽医研究所广西畜禽疫苗重点实验室;军事医学科学院军事兽医研究所;吉林大学畜牧兽医学院 |
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| 基金项目： | 广西青年科学基金(桂科青0991064);广西自然科学基金(桂科自0991219);广西科学研究与技术开发计划(桂科攻0815009-3-5) |
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| 摘 要： | 为构建猪繁殖与呼吸综合征病毒(PRRSV)重组腺病毒活载体疫苗,本研究将PRRSV GP5和M基因串联表达盒定向克隆至穿梭质粒pacAd5CMVK-NpA多克隆位点处,获得重组穿梭质粒pacAd5CMVK-NpA-GP5-M,与骨架质粒pacAd5 9.2-100共转染293AD细胞,包装出含有GP5和M基因的重组腺病毒rAd5-GP5-M。应用PCR技术、间接免疫荧光、Western blot分析,结果表明,GP5和M基因已经被成功整合到了腺病毒基因组上,且GP5和M蛋白在293AD细胞上均得到了表达;滴度测定表明,该重组腺病毒rAd5v-GP5-M的TCID50为10-6.5/mL;连续传代试验表明该重组腺病毒可在细胞内稳定繁殖,有效地转录并表达GP5和M蛋白,具有较高的遗传稳定性。该研究为PRRSV重组腺病毒疫苗的研制打下了基础。
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| 关 键 词： | 猪繁殖与呼吸综合征病毒 GP5基因 M基因 重组腺病毒 |
Construction and Expression of the Recombinant Adenovirus GP5 and M Protein of PRRSV |
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| GAO Bo,MA Ling,ZHANG Nian,MAO Yan,CHEN Feng-lian,ZHANG Shou-feng,LIU Ye,ZHANG Fei,HU Rong-liang,WU Jian-min. Construction and Expression of the Recombinant Adenovirus GP5 and M Protein of PRRSV[J]. Progress In Veterinary Medicine, 2012, 0(1): 23-27 |
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| Authors: | GAO Bo MA Ling ZHANG Nian MAO Yan CHEN Feng-lian ZHANG Shou-feng LIU Ye ZHANG Fei HU Rong-liang WU Jian-min |
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| Affiliation: | 1(1.Guangxi Veterinary Research Institute,Guangxi Key Loboratory of Animal Vaccines and New Technology, Nanning,Guangxi,530001,China;2.Veterinary Research Institute,Academy of Military Medical Sciences,Changchun, Jilin,130122,China;3.College of Animal Science and Veterinary Medicine,Jilin University,Changchun,Jilin,130062,China) |
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| Abstract: | In order to construct the recombinant vaccine of Porcine reproductive and respiratory syndrome virus(PRRSV),in this research,the ORF which contained GP5 and M genes was cloned into the shuttle vector pacAd5CMVK-NpA named pacAd5 CMVK-NpA-GP5-M and contransinfeced into 293AD cells with the backbone vector pacAd5 9.2-100.The recombinant adenovirus rAd5v-GP5-M was packed after the homologous recombination in the cells.Subsequently the recombinant virus was detected by PCR,indirect immunofluorescence test and Western blot assay.The results indicated the GP5 and M gene were intergrated into the adenovirus geneome and the proteins GP5 and M all were expressed in 293AD cells.The titre determination tests indicated the TCID50 is 10-6.5/mL.This study established the fundation for the development of recombinant adenovirus vaccine against PRRS. |
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| Keywords: | Porcine reproductive and respiratory syndrome virus GP5 protein M gene recombinant adenovirus |
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