用于质粒DNA规模化生产的大肠杆菌发酵培养基的筛选  

Selection of Liquid Media for High-density Fermentation of Recombinant E.coli for Production of Plasmids

作　　者：张泉[1] 郭晓宇[2] 袁维峰[1] 孙怀昌[1] 朱鸿飞[2] 

机构地区：[1]扬州大学兽医学院,扬州225009 [2]中国农业科学院北京畜牧兽医研究所,北京100081

出　　处：《中国生物工程杂志》2007年第1期102-105,共4页China Biotechnology

基　　金：国家"863"计划资助项目(2005AA246020)

摘　　要：为降低质粒DNA的生产成本,在标准LB培养基的基础上,利用国产试剂配制成10种大肠杆菌液体培养基,以pEGFPC3、pcDNAlacZ和pcDNKLYZ质粒转化的JM109和DH5α大肠杆菌为指示菌进行小规模发酵培养,定时采样测量OD600值及质粒产量,获得一种高性价比培养基。用该培养基培养重组大肠肝菌,绘制生长曲线,并于其对数生长中期进行42℃诱导。结果表明经42℃诱导后,重组大肠肝菌JM109和DH5α的质粒产量均有提高,重组JM109的产量比重组DH5α约提高20%,为低成本、大规模生产重组质粒提供了良好的技术保障。In order to decrease the cost of media for recombinant bacterial fermentation and plasmid DNA production, 10 different liquid media were prepared using home-made reagents and tested at small-scale for growth of recombinant E. coli. By mornitoring the bacterial density （ OD600 ）, plasmid yield and replication stability, a cost,effective medium was selected. The growth curve of pEGFPC3, pcDNA/acZ and pcDNKLYZ plasmid-harboring E. coli DH5α and JM109 were plotted in the new medium. At the mid-log growth phase, the growth temperature was shifted from 37℃ to 42℃ and cultivation was continued for 12h. Quantitative analysis showed that the plasmid yields were significantly increased, with 20% increase in JM109 than in DH5α. These data demonstrated that the new medium could be used cost-effectively for fermentation of recombinant E. coli for preparation of recombinant plasmids.

关 键 词：质粒DNA 细菌培养基 产量 

分 类 号：Q93-335[生物学—微生物学]