麻疹病毒N蛋白原核表达纯化条件的优化  

Optimization of measles virus N protein's prokaryotic expression and purification

作　　者：侯丹丹[1] 王云龙[1] 张怡青[1] 孙新城[2] 李玉林 米海 王继创 程蕾 

机构地区：[1]河南师范大学生命科学学院,河南新乡453007 [2]郑州轻工业学院食品与生物工程学院,河南郑州450001 [3]河南省生物工程技术研究中心,河南郑州450001

出　　处：《郑州轻工业学院学报（自然科学版）》2013年第6期32-34,47,共4页Journal of Zhengzhou University of Light Industry:Natural Science

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%.In order to construct a recombinant expression plasmid which induced express purification mea- sles 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 SDS- PAGE and Western blot tests showed that MV N protein molecular was 60 kD. The protein＇s purity was 90% after beinz purified hv Ni-NTA and DEAE.

关 键 词：麻疹病毒 核壳蛋白 原核表达 蛋白纯化 

分 类 号：Q786.4[生物学—分子生物学]