作　　者： ; ; ; ; ; ; ;

机构地区： 华南农业大学兽医学院

出　　处： 《华南农业大学学报》 2012年第3期393-397,共5页

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 型 蛋白 原核表达

领　　域： [农业科学] [农业科学] [农业科学]