【摘要】 通过构建重组表达质粒,诱导表达纯化麻疹病毒N蛋白.将麻疹病毒N蛋白基因片段与载体pET-32a(+)相连接,通过PCR方法扩增获得重组质粒pET-32a(+)/N,然后将重组质粒转入大肠杆菌E.coli BL21(DE3)内,并优化诱导表达时间、温度、诱导剂浓度等条件.SDS-PAGE和Western blot蛋白印迹检测表明,麻疹病毒N蛋白分子质量约为60 kD,表达产物用Ni-NTA亲和层析和DEAE纯化后,纯度达90%.更多还原

【Abstract】 In order to construct a recombinant expression plasmid which induced express purification measles virus( MV) N protein,the recombinant plamised of pET-32 a( +) / N amplified by PCR from MV N protein genes was inserted into expression vector pET-32 a( +),then it was transformed into E. coli BL21( DE3). The condition of time,temperature and concentration of IPTG were optimized. The results of SDSPAGE and Western blot tests showed that MV N protein molecular was 60 kD. The protein’ s purity was 90% after being purified by Ni-NTA and DEAE.更多还原