用大肠杆菌(Escherichia coli)表达诺如病毒P粒子  

Express Recombinant Norovirus P particles in Escherichia coli

作　　者：宋维[1] 李善爽[1] 杨桂华[1] 赵凌侠[1] 

机构地区：[1]上海交通大学农业与生物学院植物生物技术研究中心,上海200240

出　　处：《上海交通大学学报（农业科学版）》2011年第2期37-42,共6页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：国家自然科学基金项目(31071810);国家高技术研究发展计划(863计划)项目(2007AA100503)

摘　　要：为了研制番茄口服疫苗,按番茄偏爱密码子设计合成了编码诺如病毒(Norovirus,NVs)VA387毒株P粒子(P particles)基因PGENE(1 039 bp),并在该基因5'端添加了编码His-tag(6×His)的DNA序列。将构建的原核表达载体pMAL-c2-PGENE导入大肠杆菌(Escherichia coli)BL21(DE3 free),用1 mmol/L的IPTG(Isopropylβ-D-1-thiogalactopyranoside)诱导P粒子在BL21(E.coli)中表达。依据SDS-PAGE胶检测结果,对诱导温度(37℃)和时间(6 h)进行了优化,用Western blot技术证实了重组P粒子特异性和免疫活性,为研制番茄口服NVs(VA387)亚基疫苗-P粒子做了必要的准备。In order to develop edible vaccines via tomato expression system,the gene coding the VA387 Norovirus P particle（designate PGENE）（1 039 bp）was designed and synthesized according to tomato codon usage bias,and DNA sequence of the coded 6×His-tag was fused into the upstream of the PGENE for purification.Prokaryotic expression vector of the pMAL-c2-PGENE was constructed,and introduced into BL21（Escherichia coli）.The recombinant P particle protein was successfully expressed in BL21（E.coli）by 1 mmol/L IPTG（Isopropyl β-D-1-thiogalactopyranoside） induction.Based on the results of SDS-PAGE assay,the optimized induction conditions including temperature（37 ℃） and time（6 h） was established.The identity of P particle and immunocompetence were confirmed by Western blot.This research will provide indispensable preparation to develop eatable subunit vaccines through the tomato fruit expression systems.

关 键 词：诺如病毒 P粒子 原核表达系统 亚基疫苗 

分 类 号：Q78[生物学—分子生物学]