猪繁殖与呼吸综合征病毒N蛋白的原核表达、纯化与结构分析  

Purification and Structural Analysis of The Nucleocapsid of PRRS Virus Expressed from Escherichia coli

作　　者：黄金海[1] 杨汉春[1] 郭鑫[1] 陈艳红[1] 查振林[1] 

机构地区：[1]中国农业大学农业部预防兽医学重点实验室,北京100094

出　　处：《微生物学报》2004年第6期737-740,共4页Acta Microbiologica Sinica

基　　金：国家"973项目"(G19990 1190 1)~~

摘　　要：将猪繁殖与呼吸综合征病毒 (PRRSV)BJ 4毒株N基因克隆至原核表达载体pET2 8a中 ,得到重组表达载体pET2 8 N ,转化EscherichiacoliBL2 1(DE3)细胞 ,获得可溶性表达 ,表达量占菌体蛋白的 2 8%。经ProbandNi2 + 亲和层析获得重组蛋白P2 8 N ,圆二色谱 (CD)测定结果表明 ,P2 8 N重组蛋白螺旋占 2 6 1% ,折叠占 2 3 7% ,转角 19 8% ,卷曲占 30 3%。The DNA fragment encoding the nucleocapsid protein (N) of PRRSV BJ4 strain were cloned into the BamHⅠ/EcoRⅠ sites of pET28a vector to construct the expression plasmid pET28-N by designing special primers.The soluble protein (P28-N) were obtained by introducing the expression plasmid into E.coli BL21(DE3) host cell, and the amount of recombinant protein reached to 28% of the total mass of bacterial protein. PET28-N were purified by nickel-affinity column of Proband resin. The circular dichroism (CD) analysis showed that the purified PET28-N shared a significant (26%) α-helical structure, β-sheet (23.7%),β-turn (19.8%), and random coil (30.3%), respectively. Finally,the secondary stucture of N protein of PRRSV was deduced.

关 键 词：猪繁殖与呼吸综合征病毒 N蛋白 圆二色谱 二级结构 

分 类 号：S852.65[农业科学—基础兽医学]