| 凋亡素融合基因原核表达载体的构建及可溶性表达 |
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| 引用本文： | 王春艳,郝军元,张英波,王诺,张玉静.凋亡素融合基因原核表达载体的构建及可溶性表达[J].吉林农业大学学报,2007,29(3):271-274,278. |
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| 作者姓名： | 王春艳 郝军元 张英波 王诺 张玉静 |
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| 作者单位： | 吉林大学人兽共患病研究所,长春,130062 |
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| 摘 要： | 在获得凋亡蛋白融合基因的基础上,成功地构建了重组表达质粒PET-22b-SPA,将阳性重组质粒转化表达受体菌BL21(DE3)感受态细胞,经IPTG诱导表达,表达产物经聚丙烯酰胺凝胶电泳检测,凋亡蛋白融合基因获得高效表达。软件分析结果表明表达蛋白占菌体蛋白的20%,上清表达量约为10%。上清蛋白经纯化后,Western blot结果显示,利用凋亡蛋白单克隆抗体可以很好地与所表达的蛋白带特异性结合。
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| 关 键 词： | 凋亡蛋白 融合基因 可溶性表达 |
| 文章编号： | 1000-5684（2007）03-0271-04 |
| 修稿时间： | 2006-07-302006-11-01 |
Construction of Apoptin Fusion Gene Prokaryotic Expression Vector and Its Soluble Expression |
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| WANG Chun-yan,HAO Jun-yuan,ZHANG Ying-bo,WANG Nuo,ZHANG Yu-jing.Construction of Apoptin Fusion Gene Prokaryotic Expression Vector and Its Soluble Expression[J].Journal of Jilin Agricultural University,2007,29(3):271-274,278. |
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| Authors: | WANG Chun-yan HAO Jun-yuan ZHANG Ying-bo WANG Nuo ZHANG Yu-jing |
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| Institution: | Institute ofAmphixenosis, Jilin University, Changchun 130062, China |
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| Abstract: | Recombinant expression plasmid pET-22b-SPA was constructed according to recombinant plasmid.The correct recombinant expression plasmid was transformed into the host strains BL21(DE3) induced by IPTG.The specific expressed protein was detected by SDS-PAGE.The recombinant protein was expressed at high level accounting for 20% of the total bacterial protein analyzed by computer software.The amount in supernatant is about 10%.After the supernatant protein was purified,western blot showed the monoclonal antibody raised against the recombinant apoptin in murines could react to the protein expressed specifically. |
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| Keywords: | apoptin fusion gene soluble expression |
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