| 鸭圆环病毒Cap 基因在昆虫细胞杆状病毒系统中的表达及鉴定 |
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| 引用本文： | 张兴晓,邹金峰,相琪旺,王 鑫,陈 琳,谢之景,姜世金.鸭圆环病毒Cap 基因在昆虫细胞杆状病毒系统中的表达及鉴定[J].西北农业学报,2012,21(3):7-11. |
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| 作者姓名： | 张兴晓 邹金峰 相琪旺 王 鑫 陈 琳 谢之景 姜世金 |
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| 作者单位： | 1. 山东农业大学动物医学院,山东泰安,271018
2. 山东农业大学动物医学院,山东泰安 271018;山东省动物生物技术与疫病防治重点实验室,山东泰安 271018 |
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| 基金项目： | 山东省科技发展计划项目(2010GNC10914);山东省高等学校科技计划项目(J08LF07) |
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| 摘 要： | 通过PCR方法扩增完整的鸭圆环病毒Cap基因,按正确的阅读框架将其克隆入pFastBacTM HTB杆状病毒载体,将筛选的阳性重组载体pF-CP转化至含有杆状病毒穿梭载体和辅助载体的DH10Bac大肠杆菌感受态细胞,经过蓝白斑筛选和PCR鉴定,获得重组表达载体rBacmid-CP。在脂质体的介导下转染Sf9昆虫细胞,72h后获得重组杆状病毒。Western blot和间接免疫荧光(IFA)试验结果表明,表达的蛋白与原核表达Cap蛋白免疫小鼠采集的血清具有良好的反应原性,为进一步研究Cap蛋白的功能奠定基础。
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| 关 键 词： | 鸭圆环病毒 Cap基因 昆虫细胞-杆状病毒表达系统 IFA |
Expression and Identification of Cap Gene of DuCV in Bac to Bac Baculovirus Expression Systems |
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| ZHANG Xingxiao,ZOU Jinfeng,XIANG Qiwang,WANG Xin,CHEN Lin,XIE Zhijing and JIANG Shijin.Expression and Identification of Cap Gene of DuCV in Bac to Bac Baculovirus Expression Systems[J].Acta Agriculturae Boreali-occidentalis Sinica,2012,21(3):7-11. |
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| Authors: | ZHANG Xingxiao ZOU Jinfeng XIANG Qiwang WANG Xin CHEN Lin XIE Zhijing and JIANG Shijin |
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| Institution: | 1,2(1.College of Veterinary Medicine,Shandong Agricultural University,Taian Shandong 271018,China; 2.Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention,Taian Shandong 271018,China) |
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| Abstract: | The whole Cap gene of duck circovirus(DuCV) was amplified by polymerase chain reaction(PCR) and then cloned into the baculovirus donor vector pFastBacTM HTB according to the right open reading frame(ORF).The recombinant plasmids pF-CP DNA were transformed into E.coli competent cells DH10Bac containing baculovirus shuttle vector.The recombinant bacmid(rBacmid-CP) was screened and verified by PCR.The rBacmid-CP was transfected into Sf9 insect cells and the recombinant baculovirus was obtained and confirmed by Western blot and immunofluorescence assay with the antiserum was collected from the mice which were immunized with the Cap expressed in prokaryotes.The recombinant Cap would be useful resource for studying on the function of Cap. |
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| Keywords: | DuCV Cap gene Baculovirus expression vector systems IFA |
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