摘要
构建了携带asd、霍乱毒素B亚基 (CTB)基因的表达质粒 pYX2 0 1,与福氏 2a痢疾菌T32的△asd突变株FaD构成宿主 质粒平衡致死系统 ,用于在没有抗生素选择压力的情况下 ,稳定表达CTB抗原基因。以此为基础 ,构建了单独表达肠毒素性大肠杆菌CS2 6菌毛抗原基因的重组质粒 pYX2 0 2 ,以及同时表达CS6和CTB的共表达质粒 pYX2 0 3。Westernblotting和ELISA检测结果证实CS6及CTB在痢疾菌FaD中可以有效表达。重组菌免疫家兔后可诱生相应的血清抗体 ,特别是CTB的抗体效价较高 ,并持续较长时间。本研究为细菌性腹泻疫苗的研究提供了候选株。
A host\|plasmid balancing system composed with a Δ asd mutant (FaD) of an avirulent strain (T32) of Shigella flexneri 2a and plasmid harboring asd gene was used to express enterotoxigenic E.coli surface antigen 6(CS6) and V.cholerae toxin subunit B (CTB).The results of Western blotting and ELISA showed that all of recombinant plasmids (pYX201,pYX202 and pYX203) could be maintained stably and expressed CS6 and CTB respectively in T32 without any antibiotic selection.All the recombinant bacterial strains could elicit the corresponding antibodies in rabbits.The antibodies against CTB elicited by both FaD/pYX201 and FaD/pYX203 showed to be high level,and had long prolongation time,in otherwise,the antibodies against CS6 showed to be low level,indicating that higher expression level of foreign antigen may be benefit for construction of genetic multivalent vaccine.
出处
《生物工程学报》
CAS
CSCD
北大核心
2001年第1期29-34,共6页
Chinese Journal of Biotechnology
基金
军队重点基金!(96 2 5 2 2 5 )&&
