高致病性猪繁殖及呼吸综合征病毒ORF7基因的克隆及原核表达.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 20 11 2 ORF7 李国旺 苗志国 陈俊杰 (, 453003) : 猪繁殖与呼吸综合征( PRRS)是严重危害全球养猪业的重要病毒性 染病通过 PCR 方法从高致病性PRRSV WUH 1株中扩增得到结构蛋白 ORF7的基因片段, 并将目的基因插入原核表达载体 pET30a 中, 得到重组表达质粒 pET30a ORF7重组表达质粒转化大肠杆菌BL21( DE3)细胞, 经 IPTG诱导, SDSPAGE聚丙烯酰胺凝胶电泳分析表明, 融合蛋白得到 了高效表达, 表达的融合蛋白分子量约 20 D : 高致病性猪繁殖与呼吸综合征病毒 结构蛋白 ORF7基因 克隆 原核表达 融合蛋白 Cloning and Prokaryotic Expression ofGeneORF7 ofHighly Pathogenic PorcineReproductive and Respiratory Syndrome irus LiGuow ang M iao Zhiguo Chen Junjie (H enan Institute of Scienceand Technology, College of Anmi alScience,X inxiang 453003) A bstrac:t Porcine reproductive and respiratory syndrom e is an mi portan t viral infectious d isease that severely threatens global p ig industry. In this study, the gene sequence of structural protein ORF7 w as am plified from highly pathogenic PPRSV isolate WUH 1 by PCR and the target gene w as inserted into pro aryotic expression vector pET30a, generating a recom binant expression p lasm id, pET 30aORF7. The recom binant express ion p lasm idw as then used to transform com petent cells ofEscherichia coli BL21( DE3) and the ex pression w as induced by IPTG. SDSPAGE electrophoresis analysis show ed an efficient expression of the target fusion protein, of wh ich m olecularw eight w as about 20 D. K ey w ords: H ighly pathogen ic porcine reproductive and resp iratory syndrom e virus Structural protein ORF7 gene C loning Pro aryotic express ion Fusion protein ( Porcine reproductive and 15 b, 4, [ 2, 3] resp iratory syndrome, PRRS) ; PRRSV pH , ( Porcine reproductive and resp iratory syndrome ,