| 猪圆环病毒1型ORF2基因的改造及其在大肠杆菌中的表达 |
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| 引用本文： | 张福良,孙利华,宋长绪.猪圆环病毒1型ORF2基因的改造及其在大肠杆菌中的表达[J].中国动物检疫,2009,26(6):33-35. |
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| 作者姓名： | 张福良 孙利华 宋长绪 |
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| 作者单位： | 1. 安阳工学院生物与食品学院,河南安阳,455000
2. 广东省农业科院兽医研究所,广东广州,510640 |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 摘 要： | 根据GenBank中猪圆环病毒1型(PCV1)ORF2基因序列,设计合成了一对引物,对PCV1ORF2基因进行PCR扩增,将扩增出的ORF2基因(702bp)克隆入pMD18-T载体,筛选获得重组质粒命名为pMD-ORF2。由于ORF2前面有120多个信号肽和序列中有大量的稀有密码子,所以将ORF2进行改造并将改造的ORF2双酶切产物插入原核表达载体pET-32α与pET-41α,得到重组子命名为pET-ORF2-1与pET-ORF2-2。用IPTG进行诱导表达,收集菌液进行SDS-PAGE和Westernblot分析,结果表明PCV1-ORF2在pET-32α和pET-41α中获得了高效融合表达,其表达蛋白分子量约为30Kda和45Kda。
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| 关 键 词： | PCV1ORF2 克隆 改造 表达 |
Reconstruction and expression of ORF2 Gene of Porcine Cireovirus Type 1 in E.coli |
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| ZHANG Fuliang,SUN Lihu,SONG Changxu.Reconstruction and expression of ORF2 Gene of Porcine Cireovirus Type 1 in E.coli[J].China Journal Of Animal Quarantine,2009,26(6):33-35. |
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| Authors: | ZHANG Fuliang SUN Lihu SONG Changxu |
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| Institution: | 1.College of Biotechnology and Food Science, Anyang Institute of Technology, Anyang Henan 455000; 2.Veterinary Research Institute,Guangdong Academy of Agricultural Sciences,Guangzhou China 510640) |
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| Abstract: | A pair of primers was designed according to PCV1 ORF2 genome sequence in the GenBank. The ORF2 genome was cloned into pMD 18-T vector, and the recombinant plasmid was named pMD-ORF2. Then the ORF2 genome was subcloned into prokaryotic expressing vector pET-32α and pET-41α,the recombinant plasmid were named pET-ORF2-1 and pET-ORF2-2 respectively, the recombinant plasmid was induced by IPTG. The results of SDS-PAGE and Western-blot indicated that the ORF2 gene was expressed in high level, and the recombinant fusion protein was about 30Kda and 45 Kda. |
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| Keywords: | PCV1 ORF2 |
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