| 山羊痘病毒P32基因重组表达载体的构建及原核表达 |
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| 引用本文： | 程振涛,岳筠,徐春志,李永明,许乐仁,文明,周碧君.山羊痘病毒P32基因重组表达载体的构建及原核表达[J].畜牧与兽医,2007,39(12):11-13. |
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| 作者姓名： | 程振涛 岳筠 徐春志 李永明 许乐仁 文明 周碧君 |
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| 作者单位： | 贵州大学动物疫病研究所,贵州大学动物科学学院,贵州,贵阳,550025 |
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| 基金项目： | 教育部科学技术研究重点项目(206132)，贵州省优秀科技教育人才省长专项基金项目(200612)，高层次人才科研条件特助经费项目(TZJF-200606) |
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| 摘 要： | 以含山羊痘病毒贵州LD株P32基因的质粒pMD18-T-P32/LD为模板,应用PCR方法扩增P32基因,将该基因片段克隆至原核表达载体pET-28a,构建了重组表达载体pET-28a-P32/LD,转化大肠杆菌BL21(DE3)中并诱导表达。SDS-PAGE分析结果显示,有分子量约为35.5 ku的融合蛋白获得表达,与预期结果相符;Western-Blotting证实该表达蛋白具有免疫学活性。P32蛋白的成功表达为山羊痘基因工程疫苗的研制及诊断方法的建立奠定了基础。
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| 关 键 词： | 山羊痘病毒 P32基因 原核表达 |
| 文章编号： | 0529-5130（2007）12-0011-03 |
| 收稿时间： | 2007-07-09 |
| 修稿时间： | 2007年7月9日 |
Construction of recombinant expression vector with goat poxvirus P32 gene and its prokaryotic expression |
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| CHENG Zhen-tao,YUE Jun,XU Chun-zhi,LI Yong-ming,XU Le-ren,WEN Ming,ZHOU Bi-jun.Construction of recombinant expression vector with goat poxvirus P32 gene and its prokaryotic expression[J].Animal Husbandry & Veterinary Medicine,2007,39(12):11-13. |
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| Authors: | CHENG Zhen-tao YUE Jun XU Chun-zhi LI Yong-ming XU Le-ren WEN Ming ZHOU Bi-jun |
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| Abstract: | The P32 gene fragment of goat poxvirus(GPV) LD strain was amplified from pMD18-T-P32/LD by PCR,and then inserted into the pET-28a vector.The recombinant expression plasmid pET-28a-P32/LD was successfully constructed.The gene was induced by IPTG to express in Escherichia coli BL21(DE3).SDS-PAGE showed a specific protein band of 35.5 ku,which was consistent with the expected result.Western-Blotting analysis demonstrated that the specific protein band could react with antiserum of goat poxvirus,indicating its immunological activity.The experiment will provide some help for future research in GPV P32 genetic engineering vaccine and diagnostic method. |
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| Keywords: | goat poxvirus P32 gene prokaryotic expression |
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