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| 利用SUMO系统高效表达可溶性猪圆环病毒2型Cap蛋白 | |
| 引用本文： | 陈春丽,郭宇飞,陈筱薇,叶昱,王强,廖明,樊惠英.利用SUMO系统高效表达可溶性猪圆环病毒2型Cap蛋白[J].华南农业大学学报,2012,33(3):393-397. |
| 作者姓名： | 陈春丽 郭宇飞 陈筱薇 叶昱 王强 廖明 樊惠英 |
| 作者单位： | 1. 华南农业大学兽医学院,广东广州510642;四川农业大学动物医学院,四川雅安625000 2. 四川农业大学动物医学院,四川雅安,625000 3. 华南农业大学兽医学院,广东广州,510642 |
| 基金项目： | 国家自然科学青年基金，农业微生物学重点实验室开发课题，华南农业大学校长基金 |
| 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. |
| 关 键 词： | 猪圆环病毒2型 Cap蛋白 原核表达 |
| 收稿时间： | 9/6/2011 12:00:00 AM |
Efficient Expression of Cap Gene of Porcine Circovirus Type 2 by pCold-SUMO Expression Vector |
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| CHEN Chun-li , GUO Yu-fei , CHEN Xiao-wei , YE Yu , WANG Qiang , LIAO Ming , FAN Hui-ying.Efficient Expression of Cap Gene of Porcine Circovirus Type 2 by pCold-SUMO Expression Vector[J].Journal of South China Agricultural University,2012,33(3):393-397. | |
| Authors: | CHEN Chun-li GUO Yu-fei CHEN Xiao-wei YE Yu WANG Qiang LIAO Ming FAN Hui-ying |
| Institution: | 1(1 College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China; 2 College of Veterinary Medicine,Sichuan Agricultural University,Yaan 625000,China) |
| Abstract: | 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-SUMO-dCap,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 purification 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. |
| Keywords: | porcine circovirus 2 Capsid protein prokaryotic expression |
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