| 乙型脑炎病毒E基因在杆状病毒中的表达 |
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| 引用本文： | 戚龙霞,许信刚,王志昇,童德文,唐麒麟,宁蓬勃.乙型脑炎病毒E基因在杆状病毒中的表达[J].西北农业学报,2010,19(3):50-53. |
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| 作者姓名： | 戚龙霞 许信刚 王志昇 童德文 唐麒麟 宁蓬勃 |
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| 作者单位： | 西北农业科技大学,动物医学院,陕西杨陵,712100 |
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| 基金项目： | 陕西省科技攻关项目，国家大学生创新性实验计划项目，教育部新世纪优秀人才支持计划 |
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| 摘 要： | 利用PCR方法扩增日本乙型脑炎病毒(JEV)E全长基因,然后将其克隆到杆状病毒转座载体pBac-SC中,筛选重组质粒pBacSC-E,再转化入含有穿梭载体的感受态细胞DH10Bac,经抗性及蓝白斑筛选得到含E基因的重组杆状病毒DNA,以脂质体介导法将此重组杆状病毒DNA转染Sf9昆虫细胞,从而获得重组杆状病毒。用此重组杆状病毒感染Sf9昆虫细胞,经Western-blot检测表明,E基因在昆虫细胞中获得表达,表达蛋白的分子量约53 ku,与预期结果大小一致,且能与日本乙型脑炎阳性血清发生特异性反应,这为进一步研究JEV亚单位疫苗和诊断抗原奠定了基础。
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| 关 键 词： | 乙型脑炎病毒 重组杆状病毒 Sf9细胞 |
| 收稿时间： | 2009/7/13 0:00:00 |
| 修稿时间： | 2009/10/20 0:00:00 |
Expression of E Gene of Japanese Encephalitis Virus in Baculovirus System |
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| QI Longxi,XU Xingang,WANG Zhisheng,TONG Dewen,TANG Qilin and NING Pengbo.Expression of E Gene of Japanese Encephalitis Virus in Baculovirus System[J].Acta Agriculturae Boreali-occidentalis Sinica,2010,19(3):50-53. |
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| Authors: | QI Longxi XU Xingang WANG Zhisheng TONG Dewen TANG Qilin and NING Pengbo |
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| Institution: | QI Longxia,XU Xingang,WANG Zhisheng,TONG Dewen,TANG Qilin , NING Pengbo (College of Veterinary Medicine,Northwest A&F University,Yangling Shaanxi 712100,China) |
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| Abstract: | In this study,the full length E gene of Japanese encephalitis virus(JEV) was amplified by PCR and cloned into vector pBacSC to form the recombinant plasmid pBacSC-E.The recombinant pBacSC-E was identified by enzyme digestion.The pBacSC-E was transformed into E.coli strain DH10Bac.Through resistance and blue-white selection,the recombinant baculovirus was obtained from Sf9 insect cells in which recombinant baculovirus DNA transfected through the liposome-mediated.Sf9 insect cells were infected by recombinant... |
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| Keywords: | Japanese encephalitis virus Baculovirus expression Sf9 cell |
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