| 猪圆环病毒2型ORF3编码蛋白的体外表达 |
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| 引用本文： | 李增魁.猪圆环病毒2型ORF3编码蛋白的体外表达[J].畜牧与兽医,2008,40(9). |
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| 作者姓名： | 李增魁 |
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| 作者单位： | 青海大学农牧学院动物医学系,青海,西宁,810003 |
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| 摘 要： | 设计特异引物,以猪圆环病毒2型(PCV-2)杭州株HZ0201的基因组DNA为模板,PCR扩增出ORF3基因,构建了pGEX-4T-1-ORF3原核表达载体和pEGFP-C2-ORF3真核表达载体。ORF3基因全长315 bp,编码105个氨基酸。SDS-PAGE、Western blot分析及真核PK15细胞转染结果显示:ORF3蛋白在大肠杆菌中以包涵体形式存在,分子量大小约为37.7 ku;ORF3重组蛋白在真核PK15细胞的细胞核和细胞浆都有表达,尤其在细胞核中表达量较高,且对细胞有一定的毒性。
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| 关 键 词： | 猪圆环病毒2型 ORF3 原核表达 真核表达 |
In Vitro expression of ORF3 protein from porcine circovirus type 2 |
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| LI Zeng-kui.In Vitro expression of ORF3 protein from porcine circovirus type 2[J].Animal Husbandry & Veterinary Medicine,2008,40(9). |
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| Authors: | LI Zeng-kui |
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| Abstract: | The specific primers were designed and synthesized according to the published sequences of ORF3 gene of porcine circovirus type 2(PCV-2).The complete DNA fragment encoding ORF3 protein was obtained from viral DNA of PCV-2 HZ0201 by PCR and subcloned into prokaryotic expression vector pGEX-4T-1 and eukaryotic expression vector pEGFP-C2.ORF3 gene encoding 105 amino acids was 315 bp in length.The analysis of SDS-PAGE,Western blot and transfection assays showed that the ORF3 recombinant protein with the molecular weight of 37.7 ku mainly existed as inclusion body in E.coli;the recombinant protein was expressed in cellular nucleus and cytoplasma of PK-15 cells and showed some toxicity to PK-15 cells. |
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| Keywords: | porcine circovirus type 2 ORF3 prokaryotic expression eukaryotic expression |
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