| 猪圆环病毒2型ORF2-ORF1串联基因的原核表达及活性分析 |
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| 引用本文： | 闻晓波,李冬野,李树东,冉旭华,李晓娟,邵磊,王密,朴范泽,杨焕民,侯喜林,倪宏波.猪圆环病毒2型ORF2-ORF1串联基因的原核表达及活性分析[J].动物医学进展,2010,31(2):41-44. |
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| 作者姓名： | 闻晓波 李冬野 李树东 冉旭华 李晓娟 邵磊 王密 朴范泽 杨焕民 侯喜林 倪宏波 |
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| 作者单位： | 黑龙江八一农垦大学动物科技学院,预防兽医学省重点实验室,黑龙江大庆,163319 |
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| 基金项目： | 黑龙江省农垦总局课题项目(HNKXIV-08-07A);;黑龙江八一农垦大学博士科研启动项目(校启D-2007-1) |
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| 摘 要： | 为探讨原核表达的PCV-2ORF2-ORF1串联蛋白的抗原性及其作为基因工程亚单位疫苗的可能性,以构建好的PCV-2大庆分离株质粒pMD-18T/PCV-2为模板,PCR分别扩增去掉核定位信号的Cap蛋白基因(dNLS-ORF2)和Rep蛋白基因(ORF1),将其串联克隆到原核表达载体pET30a(+)上,经PCR、酶切和测序鉴定,成功构建了重组质粒pET30a-ORF2/ORF1。转化E.coli BL21(DE3)pLysS感受态细胞,经IPTG诱导后,进行SDS-PAGE电泳与Western blot分析。纯化的重组蛋白与PCV-2阳性血清发生特异性反应,并能与免疫小鼠的抗血清发生特异性反应。重组蛋白具有较好的免疫学活性,为基因工程亚单位疫苗的研制奠定了基础。
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| 关 键 词： | 猪圆环病毒2型 ORF2 ORF1 表达 |
Expression and Activity Analysis of Recombinant ORF2-ORF1 Protein of PCV-2 |
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| WEN Xiao-bo,LI Dong-ye,LI Shu-dong,RAN Xu-hua,LI Xiao-juan,SHAO Lei,WANG Mi,PIAO Fan-ze,YANG Huan-min,HOU Xi-lin,NI Hong-bo.Expression and Activity Analysis of Recombinant ORF2-ORF1 Protein of PCV-2[J].Progress In Veterinary Medicine,2010,31(2):41-44. |
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| Authors: | WEN Xiao-bo LI Dong-ye LI Shu-dong RAN Xu-hua LI Xiao-juan SHAO Lei WANG Mi PIAO Fan-ze YANG Huan-min HOU Xi-lin NI Hong-bo |
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| Institution: | Provincial Key Laboratory of Preventive Veterinary Medicine;College of Animal Science and Technology;Heilongjiang Bayi Agricultural University;Daqing;Heilongjiang;163319;China |
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| Abstract: | To analyze the antigenicity of recombinant ORF2-ORF1 protein of PCV2 and discuss the possibility as subunit vaccine. The NLS deleted ORF2 and ORF1 genes were amplified by PCR from pMD-18T/PCV-2 and cloned into the pET30a (+) vector. The recombinant plasmids pET30a-ORF2/ORF1 was successfully identified by PCR,enzymatic digestion and sequenced. The recombinant plasmid was transformed into E.coli BL21 (DE3) pLysS competent cells. Expression of recombinant protein was induced by IPTG. The expression of target p... |
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| Keywords: | Porcine circovirus 2 ORF2 ORF1 expression |
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