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构建日本血吸虫混合 DNA疫苗 ,为今后多抗原特异性 DNA疫苗的研究打下基础。在克隆 p Bluescript- Sj2 3、p Bluescript- Sj2 6、 p Bluescript- 32的基础上 ,亚克隆构建了真核表达载体 p BK- Sj2 3、 p BK- Sj2 6、 p BK- Sj32。经过酶切鉴定 ,PCR扩增鉴定及测序后 ,将此三种不同抗原质粒经不同组合后经肌肉注射免疫小鼠 ,3周后提取小鼠股四头肌DNA,特异性引物扩增获得目的基因。证实重组质粒转入了完整的日本血吸虫抗原 2 3、 2 6、 32 kd基因。且混合质粒能在肌肉组织中稳定存在。建立了日本血吸虫混合抗原 DNA疫苗 ,为以后的多特异性 DNA疫苗进一步研究奠定了基础
Construction of a mixed DNA vaccine against Schistosoma japonicum lays the foundation for the future study of multi-antigen-specific DNA vaccines. Based on the cloning of p Bluescript-Sj2 3, p Bluescript-Sj2 6, p Bluescript-32, the eukaryotic expression vectors pBK-Sj2 3, p BK-Sj2 6 and pBK-Sj32 were constructed by subcloning. After digestion identification, PCR amplification identification and sequencing, the three different antigen plasmids were immunized by intramuscular injection after different combinations, and the DNA of quadriceps femoris was extracted after 3 weeks and the specific primers were amplified for the purpose gene. Confirmed that the recombinant plasmid was transferred into the complete genome of Schistosoma japonicum antigen 2 3, 2, 6, 32 kd. And mixed plasmid can stably exist in muscle tissue. The DNA vaccine of Schistosoma japonicum mixed antigen was established, which lays the foundation for the further study of multispecific DNA vaccine