| 猪繁殖与呼吸综合征病毒Hn-1/06株GP3蛋白的原核表达与纯化 |
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| 引用本文： | 王中明,李素平,夏平安,尹彦涛,李伟娟,邱璜,刘明莉,刘玉松,崔保安.猪繁殖与呼吸综合征病毒Hn-1/06株GP3蛋白的原核表达与纯化[J].华北农学报,2009,24(1). |
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| 作者姓名： | 王中明 李素平 夏平安 尹彦涛 李伟娟 邱璜 刘明莉 刘玉松 崔保安 |
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| 作者单位： | 河南农业大学,牧医工程学院,河南,郑州,450002 |
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| 摘 要： | 根据GenBank中PRRSV美洲株ATCC VR2332的ORF3基因序列,利用Primer 5.0软件设计合成了1对特异性引物,经RT-PCR从河南分离株Hn-1/06株扩增得到了大小为765 bp的片段,并将扩增的片段插入pTG19-T载体进行测序而获得含有ORF3的阳性克隆pTG19-T-GP3.将此基因亚克隆到原核表达载体pET-32a,经筛选获得了阳性重组质粒pET-GP3,进而在IPTG的诱导下成功表达,获得了大小约为45 kDa的融合蛋白GP3-His,纯化后经Western blot检测证实表达的融合蛋白具有良好的生物学活性.从而为进一步研究PRRSV次要结构蛋白GP3的免疫特性和功能奠定了基础.
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| 关 键 词： | GP3蛋白 原核表达 |
Prokaryotic Expression of GP3 Protein of Porcine Reproductive and Respiratory Syndrome Virus Hn-1/06 Strain |
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| WANG Zhong-ming,LI Su-ping,XIA Pin-gan,YIN Yan-tao,LI Wei-juan,QIU Huang,LIU Ming-li,LIU Yu-song,CUI Bao-an.Prokaryotic Expression of GP3 Protein of Porcine Reproductive and Respiratory Syndrome Virus Hn-1/06 Strain[J].Acta Agriculturae Boreali-Sinica,2009,24(1). |
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| Authors: | WANG Zhong-ming LI Su-ping XIA Pin-gan YIN Yan-tao LI Wei-juan QIU Huang LIU Ming-li LIU Yu-song CUI Bao-an |
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| Institution: | College of Animal Husbandry and Veterinary;Henan Agricultural University;Zhengzhou 450002;China |
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| Abstract: | One pair of specific primers was designed according to the sequence of PRRSV ORF3 gene published in GenBank(Accession number PRU87392),and was used to amplify the ORF3 gene from a PRRSV Hn-1/06 strain by RT-PCR method,resulting in a 765 bp gene fragment.The RT-PCR product was directly cloned into the pTG19-T vector and the recombinant plasmid was identified by enzyme digesting and DNA sequencing.The positive clone was designated as pTG19T-GP3.After being cloned into the prokaryotic expression vector pET-32a... |
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| Keywords: | PRRSV |
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