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| 猪圆环病毒2型ORF2基因的表达及活性研究 | |
| 引用本文： | 陈涛,闫若潜,谢彩华,吴志明,张志凌,张健,刘英.猪圆环病毒2型ORF2基因的表达及活性研究[J].动物医学进展,2009,30(2). |
| 作者姓名： | 陈涛 闫若潜 谢彩华 吴志明 张志凌 张健 刘英 |
| 作者单位： | 1. 甘肃农业大学动物医学院,甘肃,兰州,730070;河南省动物疫病预防控制中心,河南郑州,450008 2. 河南省动物疫病预防控制中心,河南郑州,450008 3. 甘肃农业大学动物医学院,甘肃,兰州,730070 |
| 摘 要： | 根据猪圆环病毒2型(PCV-2)基因序列设计1对特异性引物,PCR扩增PCV-2去核定位信号(nuclear localization signal,NLS)的Cap蛋白基因(oRF2),经Bam H Ⅰ/HindⅢ双酶切后将其克隆到表达载体pQE30中并转化大肠埃希菌JM109,采用IPTG进行诱导表达,采用自行创新的纯化方法对表达的融合蛋白进行纯化,Western blot鉴定纯化后的重组Cap蛋白(rCap)活性.结果表明,成功克隆了无核定位信号的ORF2基因,其分子质量大小为579 bp;表达的rCap蛋白大小为24.1 ku左右,与预期大小相一致;经纯化后的rCap蛋白纯度在95%以上;Western blot鉴定结果表明,Cap蛋白可以和抗PCV-2抗体反应,具有很好的抗原性.本研究为进一步建立以Cap蛋白为包被抗原的PCV-2间接ELISA诊断方法以及PCV-2疫苗的研究奠定了基础. |
| 关 键 词： | 猪圆环病毒2型(PCV-2) 克隆 表达 纯化 活性 |
Study on Expression and Bioactivity of PCV-2 ORF2 Gene |
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| CHEN Tao,YAN Ruo-qian,XIE Cai-hua,WU Zhi-ming,ZHANG Zhi-ling,ZHANG Jian,LIU Ying.Study on Expression and Bioactivity of PCV-2 ORF2 Gene[J].Progress In Veterinary Medicine,2009,30(2). | |
| Authors: | CHEN Tao YAN Ruo-qian XIE Cai-hua WU Zhi-ming ZHANG Zhi-ling ZHANG Jian LIU Ying |
| Institution: | 1.College of Veterinary Medicine;Gansu Agricultural University;Lanzhou;Gansu;730070;China;2.Henan Center for Animal Diseases Control & Prevention;Zhengzhou;Henan;450008;China |
| Abstract: | The primers were designed to amplify the capsid protein gene(ORF2) of defected-nuclear localization signals(d-NLS)according to the sequence of PCV-2 ORF2 gene,and the PCR production was subsequently cloned,sequenced and sub-cloned into the prokaryotic expression vector pQE30.The expressed fusion recombinant PoIL-2(rPoIL-2) protein was purified under the process of denaturing by urea,refolding by a self-innovative renaturation buffer and dialyzing by PBS buffer.The immunological activity of the purified reco... |
| Keywords: | PCV-2 cloning expression purification bioactivity |
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