用于质粒DNA规模化生产的大肠杆菌发酵培养基的筛选

中国生物工程杂志

2007, Vol. 27

Issue (1): 102-105

研究简报

用于质粒DNA规模化生产的大肠杆菌发酵培养基的筛选

张泉¹,郭晓宇²,袁维峰²,孙怀昌²,朱鸿飞³

1. 扬州大学兽医学院
2. 信阳师范学院生命科学学院
3. 中国农业科学院

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

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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十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。

关键词： 发酵；诱导；产量

Abstract: 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, pcDNAlacZ 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 selected in this study could be used cost-effectively for fermentation of recombinant E.coli for preparation of recombinant plasmids.

Key words: Fermentation; induction; yield

收稿日期: 2006-08-24 出版日期: 2010-05-10

基金资助: 2005AA246020;国际科学基金

通讯作者: 朱鸿飞 E-mail: zqyy123@yahoo.com.cn

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引用本文:

张泉郭晓宇袁维峰孙怀昌朱鸿飞. 用于质粒DNA规模化生产的大肠杆菌发酵培养基的筛选[J]. 中国生物工程杂志, 2007, 27(1): 102-105.

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https://manu60.magtech.com.cn/biotech/CN/Y2007/V27/I1/102

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