摘要
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.
出处
《中国兽药杂志》
2006年第9期1-5,共5页
Chinese Journal of Veterinary Drug
基金
国家"863"计划资助项目(2002AA245051)
广东省重大科技专项2004A20403002)
关键词
口蹄疫病毒
衣壳前体
原核表达
泛亚株
Foot-and-Mouth disease virus
capsid precursor
prokaryotic expression