| 柔嫩艾美耳球虫F2杂交株Rhomboid基因在大肠杆菌中的表达 |
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| 引用本文： | 张广海,李建华,宫鹏涛,张西臣,张国才. 柔嫩艾美耳球虫F2杂交株Rhomboid基因在大肠杆菌中的表达[J]. 吉林农业大学学报, 2009, 31(1) |
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| 作者姓名： | 张广海 李建华 宫鹏涛 张西臣 张国才 |
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| 作者单位： | 吉林大学畜牧兽医学院,长春,130062;吉林大学畜牧兽医学院,长春,130062;吉林大学畜牧兽医学院,长春,130062;吉林大学畜牧兽医学院,长春,130062;吉林大学畜牧兽医学院,长春,130062 |
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| 基金项目： | 国家自然科学基会项目 |
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| 摘 要： | 根据ET-Rhomboid基因开放阅读框,插入有利于表达的酶切位点设计引物,以虫体DNA为模板扩增并克隆了ET-Rhomboid基因,并将该基因与PET-28a(+)质粒载体连接,构建成了原核表达载体PET-ET-Rhomboid.将构建好的表达质粒转入大肠杆菌DE3,经IFTG诱导表达后,用SDS-PAGE检测表达产物,用蛋白印迹(Western blotting)检测其反应原性.结果表明:含重组质粒的工程菌有明显的表达产物为31 kD的融合蛋白,与推测的融合蛋白的分子量吻合,蛋白印迹检测具有反应原性.
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| 关 键 词： | 柔嫩艾美耳球虫 Rhomboid基因 原核表达 |
Expression of Rhomboid Gene of Eimeria tenella F2 Strain in E. Coli |
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| ZHANG Guang-hai,LI Jian-hua,GONG Peng-tao,ZHANG Xi-chen,ZHANG Guo-cai. Expression of Rhomboid Gene of Eimeria tenella F2 Strain in E. Coli[J]. Journal of Jilin Agricultural University, 2009, 31(1) |
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| Authors: | ZHANG Guang-hai LI Jian-hua GONG Peng-tao ZHANG Xi-chen ZHANG Guo-cai |
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| Affiliation: | College of Animal Science and Veterinary Medicine;Jilin University;Changchun 130062;China |
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| Abstract: | Based on the ORF and restriction sites of the ET-Rhomboid,ET-Rhomboid gene was amplified and cloned.Connecting the ET-Rhomboid with the plasmid vector PET-28a(+),prokaryotic expression vector PET-ET-Rhomboid was constructed.The PET-ET-Rhomboid was transformed into E.coli DE3 and induced by IPTG and then detected by SDS-PAGE and Western blotting.The results indicated that the recombinant bacteria had obvious expression product which was approximately 31 kD and had an reactivity detected by Western blotting. |
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| Keywords: | Eimeria tenella Rhomboid gene prokaryotic expression |
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