口蹄疫病毒泛亚株衣壳蛋白在大肠杆菌中高效表达和鉴定

Fusion Expression of the Entire Capsid Precursor (P1) Gene of the Foot-and-Mouth Disease Virus in Escherichia coli

以pT-P1质粒为模板,应用引物对P1-S2/P1-A2进行PCR扩增,获得口蹄疫病毒（FMDV）泛亚株全衣壳前体基因（P1）片段.用EcoRI 酶切并连接将P1克隆入表达载体pSOC中,以引物对Pr.78/P1-A2进行PCR扩增,鉴定P1基因插入方向正确.在大肠杆菌中以P1-SOC融合蛋白（约98ku）的形式获得高效表达（21%）.在0.5～3 mmol/L IPTG诱导浓度和37 ℃220 r/m振摇培养1～4 h条件下表达量保持稳定.表达产物经T4-3C蛋白酶作用,裂解后可产生多种蛋白.出现与抗口蹄疫血清反应的蛋白条带.

The entire capsid precursor gene （ P1 ） of the Foot-and-Mouth Disease virus （FMDV） was cloned into the expression vehicle （pSOC）, and high expressivity （21%） of it in the form of P1-SOC fusion protein （about 98 ku） was attained in E. coli. The expression level remained constant when 0.5 - 3 mmol/L IPTG was used for induction and incubation at 37 ℃ for 1 -4 h was done on a orbital shaker running at 220 r/m. After cleavage with T4-3C protease, the expression product produced several protein bands that could react with FMD positive immune serum.