title: Effects of medium composition on the production of plasmid DNA vector potentially for human gene therapy.
abstract: Plasmid vector is increasingly applied to gene therapy or gene vaccine. The production of plasmid pCMV-AP3 for cancer gene therapy was conducted in a modified MBL medium using a recombinant E. coli BL21 system. The effects of different MMBL components on plasmid yield, cell mass and specific plasmid DNA productivity were evaluated on shake-flask scale. The results showed that glucose was the optimal carbon source. High plasmid yield (58.3 mg/L) was obtained when 5.0 g/L glucose was added to MMBL. Glycerol could be chosen as a complementary carbon source because of the highest specific plasmid productivity (37.9 mg DNA/g DCW). After tests of different levels of nitrogen source and inorganic phosphate, a modified MMBL medium was formulated for optimal plasmid production. Further study showed that the initial acetate addition (less than 4.0 g/L) in MMBL improved plasmid production significantly, although it inhibited cell growth. The results will be useful for large-scale plasmid production using recombinant E. coli system.
authors.affiliation: Institute Bioengineering, School of Material Science and Chemical Engineering, Zhejiang University, Hangzhou 310027, China. znxu@zju.edu.cn
authors.forename: Zhi-Nan
authors.initials: ZN
authors.lastname: Xu
authors.forename: Wen-He
authors.initials: WH
authors.lastname: Shen
authors.forename: Hao
authors.initials: H
authors.lastname: Chen
authors.forename: Pei-Lin
authors.initials: PL
authors.lastname: Cen
mesh_terms: D000085:Acetates / Q000494:pharmacology; D002244:Carbon / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D003470:Culture Media / Q000737:chemistry* / Q000494:pharmacology*; D004247:DNA / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000166:cytology / Q000187:drug effects* / Q000235:genetics*; D015316:Genetic Therapy / Q000295:instrumentation*; D005822:Genetic Vectors / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D009584:Nitrogen / Q000494:pharmacology; D010710:Phosphates / Q000494:pharmacology; D010957:Plasmids / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000085:Acetates; D003470:Culture Media; D010710:Phosphates; D002244:Carbon; D004247:DNA; D009584:Nitrogen