【摘要】 根据马动脉炎病毒膜蛋白(M)的核苷酸序列,设计合成了 1 对引物,从 pUC 18 M质粒中扩增出截短的膜蛋白M基因片段。对该片段及 pGEX 6P 1 载体双酶切并连接,成功构建了原核表达载体pGEX 6P Mt。将此重组质粒转化BL21(DE3)宿主菌,对培养条件及诱导表达条件等影响表达的因素进行了优化;诱导后菌体裂解物经 SDS PAGE分析发现,在约 34 ku处出现了 1条特异性的蛋白条带,其分子质量与预期的M截短蛋白的分子质量相符,并随着诱导时间的延长而变化,在诱导后4 h达到高峰。结果表明,膜蛋白 M基因截短型在大肠埃希氏菌中得到了高效表达。更多还原

【Abstract】 The gene of truncated membrane protein of equine arteritis virus was ampilified from pUC18-M plasmid with equine arteritis virus ORF6 sequence by PCR. The PCR product was sequenced, purified and digested with EcoRⅠ and XhoⅠ, then directly cloned into the prokaryotic expression vector pGEX-6P-1. Consequently the recombinant plasmid was constructed, which was designated as pGEX-6P-Mt. pGEX-6P-Mt was transformed into the host cell BL21(DE3) and the expression procedure was optimized. The result showed that the nucleocapsid protein can be expressed efficiently with 0.8mmol/L IPTG and (4-hour-)induction.更多还原