| 山羊痘病毒P32基因在杆状病毒中的表达 |
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| 引用本文： | 郭建刚,郑敏,刘棋,陆承平,李华明,邓朝阳,邹联斌,姚火春.山羊痘病毒P32基因在杆状病毒中的表达[J].中国兽医科技,2005,35(12):950-953. |
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| 作者姓名： | 郭建刚 郑敏 刘棋 陆承平 李华明 邓朝阳 邹联斌 姚火春 |
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| 作者单位： | [1]南京农业大学动物医学院,江苏南京210095 [2]广西壮族自治区兽医防疫检疫站,广西南宁530001 |
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| 摘 要： | 采用PCR亚克隆了山羊痘病毒P32基因片段,将其插入杆状病毒转座载体质粒pFastBacⅠ中,构建了重组质粒pFastBac-P32;再将该质粒转化DH10 Bac感受态细菌,进行体内重组,然后经三重抗性及蓝白斑筛选,获得杆状病毒重组质粒Bacmid-P32;将Bacmid-P32转染Sf9细胞,获得重组杆状病毒;最后用SDS-PAGE和Western-blotting对P32基因的表达进行检测.结果表明,P32基因在重组杆状病毒中获得表达,且表达产物可以被多抗所识别.
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| 关 键 词： | 山羊痘病毒 P32基因 杆状病毒 表达 |
| 文章编号： | 1000-6419（2005）12-0950-04 |
| 收稿时间： | 2005-08-29 |
Expression of P32 gene of goatpox virus in baculovirus |
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| Institution: | 1. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095 ,China ; 2. Guangxi General Veterinary Agency, Nannin 530001 ,China |
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| Abstract: | The P32 gene of goatpox virus was amplified by PCR, and then cloned into a baculovirus expression vector pFastBac I . A recombinant plasmid pFastBac-P32 was constructed and then transformed into DH10Bac competent cells. White colonies of the recombinant Bacmid-P32 were obtained by screening with three antibiotics and selection of blue-white patches. After the Bacmid-P32 was transfected into Sf9 insect cells, the recombinant baculovirus was obtained by three passages. Finally, the expressed product from the P32 gene was detected by SDS-PAGE and Western-blotting. Result showed that the P32 gene was expressed successfully in |
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| Keywords: | goatpox the recombinant baculovirus virus P32 gene baculovirus expression |
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