| 羊痘病毒P32基因的克隆与原核表达 |
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| 引用本文： | 陈轶霞,郑亚东,窦永喜,乔军,景志忠,才学鹏.羊痘病毒P32基因的克隆与原核表达[J].动物医学进展,2007,28(1):31-34. |
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| 作者姓名： | 陈轶霞 郑亚东 窦永喜 乔军 景志忠 才学鹏 |
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| 作者单位： | 1. 甘肃农业大学动物医学院,甘肃兰州,730070;中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,甘肃省动物寄生虫病重点实验室,甘肃兰州,730046
2. 中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,甘肃省动物寄生虫病重点实验室,甘肃兰州,730046 |
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| 摘 要： | 根据发表的羊痘病毒P32基因,设计一对特异性引物,PCR扩增P32全长基因,将其克隆入pMD-18-T载体,获得重组质粒pMD-P32.再将P32基因亚克隆入原核表达载体pGEX-4T-1,获得重组表达质粒pGEX-P32,转化大肠埃希菌BL21后用IPTG诱导,表达产物经SDS-PAGE分析.结果表明,P32全长基因在大肠埃希菌中以融合形式成功表达,分子质量为58 ku左右,与预期大小相符.
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| 关 键 词： | 羊痘病毒 P32基因 克隆 表达 |
| 文章编号： | 1007-5038（2007）01-0031-04 |
| 收稿时间： | 2006-09-30 |
| 修稿时间： | 2006年9月30日 |
Cloning of the P32 Gene of Capripoxvirus and Construction of Its Prokaryotic Expression Vector |
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| CHEN Yi-xia,ZHEN Ya-dong,DOU Yong-xi,QIAO Jun,JING Zhi-zhong,CAI Xue-peng.Cloning of the P32 Gene of Capripoxvirus and Construction of Its Prokaryotic Expression Vector[J].Progress In Veterinary Medicine,2007,28(1):31-34. |
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| Authors: | CHEN Yi-xia ZHEN Ya-dong DOU Yong-xi QIAO Jun JING Zhi-zhong CAI Xue-peng |
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| Abstract: | A pair of PCR primes were designed according to the published sequence of capripoxvirus major antigen p32. The P32 gene was multiplied by PCR and cloned into the pMD-18-T vector,we got the recombinant bacmid pMD-p32. Then the P32 gene was cloned into the E. coli expression vector pGEX-4T-1 and transformed into BL21 cells. After sequencing the gene,we got the recombinant plasmid pGEX-p32. After induction by IPTG in E. coli BL21, the 58 ku recombinant protein was obtained by SDS-PAGE. |
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| Keywords: | Capripoxvirus P32 gene clonie expression |
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