Effects of medium composition on the production of plasmid DNA vector potentially for human gene therapy.
- Author:
Zhi-Nan XU
1
;
Wen-He SHEN
;
Hao CHEN
;
Pei-Lin CEN
Author Information
1. Institute Bioengineering, School of Material Science and Chemical Engineering, Zhejiang University, Hangzhou 310027, China. znxu@zju.edu.cn
- Publication Type:Journal Article
- MeSH:
Acetates;
pharmacology;
Carbon;
pharmacology;
Cell Division;
drug effects;
Culture Media;
chemistry;
pharmacology;
DNA;
biosynthesis;
genetics;
Escherichia coli;
cytology;
drug effects;
genetics;
Genetic Therapy;
instrumentation;
Genetic Vectors;
biosynthesis;
genetics;
Humans;
Nitrogen;
pharmacology;
Phosphates;
pharmacology;
Plasmids;
biosynthesis;
genetics
- From:
Journal of Zhejiang University. Science. B
2005;6(5):396-400
- CountryChina
- Language:English
-
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.
