| 鸡新城疫病毒La Sota株F基因克隆及原核表达 |
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| 引用本文： | 符芳,姜北宇,张莉,高轩,张飚.鸡新城疫病毒La Sota株F基因克隆及原核表达[J].华北农学报,2006,21(3):117-120. |
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| 作者姓名： | 符芳 姜北宇 张莉 高轩 张飚 |
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| 作者单位： | 1. 河北农业大学,动物科技学院,河北,保定,071001
2. 北京市农林科学院畜牧兽医研究所,北京,100089 |
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| 摘 要： | 参照NDVLa Sota株核酸序列(AF077761)设计1对引物,利用RT-PCR扩增F基因并得到了长为1 700 bp的片段,将其克隆到pGEM-T easy vector中,经酶切鉴定和测序后克隆进原核表达载体pET-32a,将重组表达质粒转化BL21(DE3),在IPTG诱导下表达约83 kd的融合蛋白,SDS-PAGE电泳和Western blotting检测证实该基因片段获得高效表达且表达产物具有免疫学活性。
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| 关 键 词： | 新城疫病毒 F基因 克隆 原核表达 |
| 文章编号： | 1000-7091（2006）03-0117-04 |
| 收稿时间： | 2005-12-26 |
| 修稿时间： | 2005-12-26 |
Cloning and Prokaryotic Expression of the F Gene of NDV La Sota Strain |
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| FU Fang,JIANG Bei-yu,ZHANG Li,GAO Xuan,ZHANG Biao.Cloning and Prokaryotic Expression of the F Gene of NDV La Sota Strain[J].Acta Agriculturae Boreali-Sinica,2006,21(3):117-120. |
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| Authors: | FU Fang JIANG Bei-yu ZHANG Li GAO Xuan ZHANG Biao |
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| Institution: | 1. College of Animal Science and Veterinary Medicine, Hebei Agricultural University, Baoding 071001, China; 2. Institute of Animal Science and Veterinary Medicine, Beijing Municipal Academy of Agricultural and Forestry Science, Beijing 100089, China |
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| Abstract: | A pair of primers was designed according to the sequence of F gene of NDV La Sotastrain(AF077761) for amplifying the F gene by RT-PCR.1 700 bp DNA fragment was amplified and cloned into pGEM-T easy vector.After restriction endonuclease analysis and sequencing,the fragment was inserted into the expression vector pET-32a.The recombinant plasmid produced a 83 kd fusion protein in E.coli BL21(DE3) under the induction of IPTG.The expression quantity and immunoreactivity of the fusion protein was detected by SDS-PAGE electrophoresis and Western blotting. |
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| Keywords: | Newcastal disease virse(NDV) F gene Clone Prokaryotic expression |
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