仙台病毒核衣壳蛋白基因的克隆与原核表达  

Cloning and Expression of Nucleoprotein Gene of Sendai Virus in Prokaryotic System

作　　者：姜骞[1] 刘怀然[1] 张龄[1] 朱洪伟[2] 曲连东[1] 

机构地区：[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

出　　处：《中国实验动物学报》2006年第2期142-144,共3页Acta Laboratorium Animalis Scientia Sinica

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。Objective To induce expression of nucleoprotein gene of Sendai virus （SV） in prokaryotic system. Methods A 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 （ DE3 ）pLysS was induced with 1 mmol/L IPTG. Results SDS-PAGE showed that the recombinant protein was about 60 Ku which could react with rabbit-anti Sendai virus serum in Western blot assay. Conclusion The recombinant nucleoprotein as an antigen may provide a basis for diagnosis of Sendal virus.

关 键 词：仙台病毒 基因 克隆 基因表达 

分 类 号：R373-33[医药卫生—病原生物学]