摘要
目的:利用真核表达质粒pRSC,构建结核杆菌抗原85A(Ag85A)与小鼠白细胞介素21(mIL21)共表达重组体pRSC-mIL21-Ag85A,为研究新型结核杆菌DNA疫苗提供新的策略。方法:从质粒pcDNA3.1-mIL21中经PCR扩增出mIL21基因,并插入质粒pRSC中构成pRSC-mIL21;再从pIRES-Ag85A质粒中经PCR扩增出Ag85A基因,构建于pRSC-mIL21重组质粒上,成为共表达DNA疫苗pRSC-mIL21-Ag85A。结果:经酶切、基因测序证实,该疫苗构建正确并能成功表达目的基因。共表达DNA疫苗免疫小鼠后,CTL活性、特异性淋巴细胞增殖水平及小鼠血清特异性抗体均呈有意义的提高。结论:结核杆菌Ag85A与mIL21共表达DNA疫苗能诱导小鼠免疫反应,为进一步研究DNA疫苗抗结核杆菌攻击的免疫防护效应奠定了基础。
Objective： To construct an eukaryotic co-expression recombinant containing mouse interleukin 21（IL-21） and tuberculosis antigen 85A （Ag85A）, it provides the new strategy for further development of new type anti-tuberculosis DNA vaccines. Methods： The gene of mIL-21 was amplified from plasmid pcDNA3.1-mIL21 by PCR and cloned directly into the plasmid pRSC, forming recombinant pRSC-mIL21. The gene of Ag85A was amplified from the plasmid pIRES-Ag85A by PCR and cloned directly into the recombinant pRSC-mIL21 again, forming co-expression DNA vaccine pRSC-mIL21-Ag85A finally. Results： It was identified by the endonuclease digestion and DNA sequencing. The results showed that the co-expression DNA vaccine had been constructed successfully and the target gene was expressed correctly. When mice were immunized with the pRSC-mIL21-Ag85A, the CTL activities, the spleen cell proliferation responses to Ag85A and the antibody fiter of serum were significantly increased respectively compared with the control mice immunized with blank plasmid. Conclusion： The eukaryotic co-expression DNA vaccine pRSC-mIL21-Ag85A can induce immune responses in mice. This study has laid a foundation for further research about DNA vaccine protective function in immunized mice challenged by Mycobacterium tuberculosis.
出处
《生物技术通讯》
CAS
2007年第1期25-28,共4页
Letters in Biotechnology
基金
东南大学校基金项目(92230021991)
