中国寄生虫学与寄生虫病杂志 ›› 2001, Vol. 19 ›› Issue (4): 4-208.

• 论著 • 上一篇    下一篇

日本血吸虫多价DNA疫苗的构建

李柳哲;石佑恩;江侃
  

  1. 华中科技大学同济医学院寄生虫学教研室!武汉430030
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2001-08-30 发布日期:2001-08-30

Construction of Multivalent DNA Vaccine against Schistosoma japonicum

LI Liu-zhe;SHI You-en;JIAGN Kan   

  1. Department of Parasitology;Tongji Medical College;Huazhong University of Science and Technology;Wuhan 430030
  • Received:1900-01-01 Revised:1900-01-01 Online:2001-08-30 Published:2001-08-30

摘要:   目的 构建日本血吸虫多价 DNA疫苗。 方法 在本室已构建的克隆载体 p Bluescript- Sj2 6、p Bluescript- Sj32及 p Bluescript- Sj2 3的基础上 ,根据 Sj2 6 k Da、Sj32 k Da和 Sj2 3k Da基因序列分别设计一对引物 ,且在引物中引入 1个含多肽接头甘氨酸 -甘氨酸 -丝氨酸 (G- G- S)的碱基序列。首先将 Sj2 3基因连接入载体 p BK中 ,构建重组质粒 p BK - Sj2 3,然后将 Sj2 6 k Da或 32 k Da基因分别连接入载体 p BK- Sj2 3中 ,构建成多价 DNA疫苗p BK- Sj2 6 - Sj2 3和 p BK- Sj32 - Sj2 3。对重组基因鉴定后 ,检测了质粒在小鼠肌肉中的表达情况。 结果 多价 DNA疫苗 p BK- Sj2 6 - Sj2 3和 p BK- Sj32 - Sj2 3转入了完整的 Sj2 6、Sj2 3和 Sj32基因。多价基因质粒能在小鼠肌肉组织中表达。结论 构建了日本血吸虫多价 DNA疫苗。

关键词: 日本血吸虫, Sj26, Sj23, Sj32, 多价, DNA疫苗

Abstract:  Objective To construct a multivalent DNA vaccine.\ Methods\ The multivalent DNA vaccine candidates pBK Sj26 Sj23,pBK Sj32 Sj23 were constructed based on the plasmids pBluescript Sj26,pBluescript Sj32 and pBluescript Sj23 with three pairs of specific primers using DNA recombinant technique. In the primers, a synthetic linker sequence encoding a peptide was designed,and the antigen genes Sj26 and Sj23,Sj32 and Sj23 were then ligated. After identification, the quadriceps muscle of mice were immunized with the multivalent antigen genes. Four weeks after immunization, the multivalent antigen genes were present in the muscular tissue of mice by PCR.\ Results\ The eukaryotic plasmids including multivalent antigens of S.japonicum were constructed successfully, and the plasmids including multivalent antigen gene could be stably existing in the muscle tissue of mice and the multivalent antigens could be expressed in the muscle tissue cells of mice.\ Conclusion\ A multivalent S.japonicum DNA vaccine has been established.

Key words: Schistosoma japonicum, Sj26, Sj23, Sj32, multivalent, DNA vaccine