| 仙台病毒核衣壳蛋白基因的克隆与原核表达 |
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| 引用本文： | 姜骞,刘怀然,张龄,朱洪伟,曲连东. 仙台病毒核衣壳蛋白基因的克隆与原核表达[J]. 中国实验动物学报, 2006, 14(2): 142-144 |
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| 作者姓名： | 姜骞 刘怀然 张龄 朱洪伟 曲连东 |
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| 作者单位： | 1. 中国农科院哈尔滨兽医研究所,哈尔滨,150001
2. 八一农垦大学,大庆,150030 |
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| 摘 要： | 目的利用原核表达系统表达仙台病毒(Sendai Virus,SV)核衣壳蛋白。方法根据GenBank上发表的仙台病毒(Sendai Virus,SV)核衣壳蛋白基因序列,设计并合成一对引物,通过RT-PCR扩增出核衣壳蛋白全长cDNA序列,将其克隆至原核表达载体pET-30a,转化大肠杆菌BL21(DE3)PlysS,于37℃1mmol/LIPTG条件下诱导表达,大肠杆菌裂解物经SDS-PAGE分析,在相对分子质量约60×103处出现一新蛋白带,与预期目的蛋白分子量相符。结果Western blot检测表明,表达产物能与兔抗SV阳性血清发生特异性反应,出现单一反应带,表明其具有免疫原性。结论为建立以重组NP蛋白为诊断抗原检测SV奠定基础。
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| 关 键 词： | 仙台病毒 基因 克隆 基因表达 |
| 文章编号： | 1005-4847(2006)02-0142-03 |
| 修稿时间： | 2005-02-03 |
Cloning and Expression of Nucleoprotein Gene of Sendai Virus in Prokaryotic System |
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| JIANG Qian,LIU Huai-ran,ZHANG Ling,ZHU Hong-wei,QU Lian-dong. Cloning and Expression of Nucleoprotein Gene of Sendai Virus in Prokaryotic System[J]. Acta Laboratorium Animalis Scientia Sinica, 2006, 14(2): 142-144 |
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| Authors: | JIANG Qian LIU Huai-ran ZHANG Ling ZHU Hong-wei QU Lian-dong |
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| Abstract: | Objective To induce expression of nucleoprotein gene of Sendai virus (SV) in prokaryotic system. MethodsA pair of primers were designed and synthesized according to the SV nucleoprotein gene sequence published by GenBank. Nucleoprotein gene was amplified by RT-PCR. Recombinant pET-SN was constructed after the SN gene was sequenced. BL21(DE_3)pLysS was induced with 1 mmol/L IPTG. ResultsSDS-PAGE showed that the recombinant protein was about 60 Ku which could react with rabbit-anti Sendai virus serum in Western blot assay. ConclusionThe recombinant nucleoprotein as an antigen may provide a basis for diagnosis of Sendai virus. |
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| Keywords: | Sendai Virus Gene Cloning Gene Expression |
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