摘要
目的 较高效率分泌表达大肠杆菌不耐热肠毒素B亚单位并鉴定其性质。方法 将编码成熟大肠杆菌不耐热肠毒素 (LTB)的基因引入pET2 2b(+)并转化E .coliBL2 1(DE3) ;重组工程菌在改良M9 CAA培养基内进行表达 ;对分泌表达的LTB进行纯化后进行神经节苷脂 (GM1)结合活性、免疫原性与黏膜免疫佐剂活性的鉴定。结果 LTB既能分泌表达至胞周质又能以包含体形式表达 ,两者都与pelB信号肽准确分离。纯化出的LTB具有正常的GM1结合活性、免疫原性及黏膜免疫佐剂活性。结论 pET2 2b(+)质粒的pelB信号肽可使LTB以较高效率分泌表达。
Objective To efficiently express the heat-labile enterotoxin B subunit (LTB) in a secretory form and to identify its biological character. Methods The gene encoding mature LTB was inserted into expression vector pET22b(+) and then the recombinant plasmid was transformed in E.coli BL21 (DE3). The recombinant bacteria were expressed in modified M9-CAA cultural media. The GM1 binding activity, immunogenicity, and mucosal immunoadjuvanticity of the recombinant LTB were identified after purification. Results The recombinant LTB could expressed in a secretory form in periplasm as well as in a form of inclusion body. Mature recombinant LTB was separated from pelB signal peptide exactly. The purified LTB had normal GM1 binding activity, immunogenicity, and mucosal immunoadjuvanticity. Conclu- sion LTB can be efficiently expressed in secretory form under introduction of pelB signal peptide of plasmid pET22b(+).
出处
《免疫学杂志》
CAS
CSCD
北大核心
2004年第5期364-368,共5页
Immunological Journal
基金
国家重点科技计划项目 (96 90 1 0 1 54)
"863"生物与现代农业技术领域生物工程技术主题课题 (2 0 0 1AA2 1 51 61 )资助
