利用SUMO系统高效表达可溶性猪圆环病毒2型Cap蛋白  

Efficient Expression of Cap Gene of Porcine Circovirus Type 2 by pCold-SUMO Expression Vector

作　　者：陈春丽[1,2] 郭宇飞[2] 陈筱薇[1] 叶昱[1] 王强[1] 廖明[1] 樊惠英[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]四川农业大学动物医学院,四川雅安625000

出　　处：《华南农业大学学报》2012年第3期393-397,共5页Journal of South China Agricultural University

基　　金：国家自然科学青年基金(30800826);农业微生物学重点实验室开发课题(20090010);华南农业大学校长基金(5500-K08240)

摘　　要：利用pCold-SUMO可溶性原核表达载体,构建表达猪圆环病毒2型缺失核定位信号肽的Cap基因的重组表达载体pCold-SUMO-dCap,并将其转入Arctic-Express表达菌株中15℃低温诱导表达.表达产物经SDS-PAGE分析,结果表明SUMO-dCap融合蛋白获得了高效表达,可溶性SUMO-dCap融合蛋白约占总融合蛋白的50%;用NI-NTA树脂纯化可溶性的融合蛋白,然后利用SUMO蛋白酶特异性去除SUMO标签,从而得到不含任何标签的dCap蛋白,进一步的Western-blot分析表明,表达的dCap蛋白具有良好的反应原性.The objective of this study was to utilize pCold-SUMO expression vector to efficiently express capsid （Cap）protein gene without nuclear localization signal （NLS） of porcine circovirus 2 （PCV2）. Cap gene without NLS was subcloned into the pCold-SUMO expression vetor, which named as pCold-SUMOdCap, and the recombinant plasmid was transformed into Arctic-Express competent cells. It was induced by IPTG at 15℃ and efficiently produced active fusion protein of SUMO-dCap. SDS-PAGE analysis of the recombinant protein demonstrated that soluble SUMO-dCap in the supernatant was expressed at approximately 50% of total recombinant fusion proteins. Then, the soluble SUMO-dCap was purified by Ni-NTA resin purification kits, whereas the SUMO tag was removed by SUMO protease. In addition,the puri- fication effect and specificity of recombinant fusion protein and capsid protein without NLS were detected by Western-blot assay. The results showed that both of them had been well purified and possessed good reactionogenicity.

关 键 词：猪圆环病毒2型 CAP蛋白 原核表达 

分 类 号：S852.65[农业科学—基础兽医学]