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目的以乳酸乳球菌(Lactococcus lactis)为载体,将鼠疫抗原LcrV基因导入乳酸乳球菌内,构建重组肠道微生态菌株,作为黏膜免疫疫苗的先期探索和尝试。方法采用酸诱导P170启动子,乳酸乳球菌本身的SP310mut2信号肽,将鼠疫杆菌LcrV抗原结构基因克隆到质粒pAM J397上,电转化感受态Lactococcus lactis PSM565。结果经重组子PCR鉴定,SDS-PAGE检测,W estern-b lot鉴定,在Lactococcus lactisPSM565/pAM J397-V培养基上清中获得了38 kD的鼠疫抗原LcrV蛋白。结论在乳酸乳球菌中成功表达了鼠疫V抗原,为下一步鼠疫黏膜疫苗的研制打下基础。
Objective To introduce Lactococcus lactis into Lactococcus lactis by using Lactococcus lactis as a carrier, and to construct recombinant gut microbiological strains as an early exploration and attempt to immunize mucosal vaccine. Methods The P170 promoter and the SP310mut2 signal peptide of Lactococcus lactis were induced by acid. The structural gene of Lactococcus lactis LcrV antigen was cloned into plasmid pAM J397 and transformed into competent Lactococcus lactis PSM565. Results The 38 kD plague antigen LcrV protein was obtained in the supernatant of Lactococcus lactisPSM565 / pAM J397-V by recombinant PCR, SDS-PAGE and W estern-b lot. Conclusion The plague V antigen was successfully expressed in Lactococcus lactis, laying the foundation for the further development of the plague vaccine.