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目的 :高密度、高质粒拷贝数培养大肠杆菌生产重组 DNA疫苗 pc D- fla B。 方法 :应用 FU S- 5 L自控发酵罐 ,采用补料分批培养技术 ,控制补料的补加使溶氧控制在 30 %～ 6 0 % ,p H控制在 7左右 ,培养中后期采用 4 2℃培养以提高质粒拷贝数。 结果 :重组菌 E.coli DH5α/ pc DNA 3- fla B发酵液光密度 (D6 0 0 )达到 4 5。经纯化后最终质粒 DNA得率为 5 0 m g/ L。质粒没有发生丢失现象。 结论 :本实验为钩端螺旋体 DNA疫苗的大规模生产奠定了基础。
OBJECTIVE: To produce Escherichia coli producing recombinant DNA vaccine pc D-fla B with high density and high plasmid copy number. Methods: FU-5 L self-control fermenter was used. The fed-batch culture technique was used to control the addition of supplements to control the dissolved oxygen concentration at 30% ~ 60%, p H was controlled at about 7 and 4 Incubate at 2 ° C to increase plasmid copy number. Results: The optical density (D6 0 0) of the recombinant E. coli DH5α / pc DNA 3-fla B fermentation broth reached 45. The final plasmid DNA yield was 50 mg / L after purification. No loss of plasmid occurs. Conclusion: This experiment lays the foundation for large-scale production of Leptospira DNA vaccine.