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Journal of Zhejiang University SCIENCE B 2005 Vol.6 No.5 P.396~400


Effects of medium composition on the production of plasmid DNA vector potentially for human gene therapy

Author(s):  XU Zhi-nan, SHEN Wen-he, CHEN Hao, CEN Pei-lin

Affiliation(s):  Institute Bioengineering, School of Material Science and Chemical Engineering, Zhejiang University, Hangzhou 310027, China

Corresponding email(s):   znxu@zju.edu.cn

Key Words:  Plasmid DNA, Growth medium, Gene therapy

XU Zhi-nan, SHEN Wen-he, CHEN Hao, CEN Pei-lin. Effects of medium composition on the production of plasmid DNA vector potentially for human gene therapy[J]. Journal of Zhejiang University Science B, 2005, 6(5): 396~400.

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author="XU Zhi-nan, SHEN Wen-he, CHEN Hao, CEN Pei-lin",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

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%T Effects of medium composition on the production of plasmid DNA vector potentially for human gene therapy
%A XU Zhi-nan
%A SHEN Wen-he
%A CEN Pei-lin
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%DOI 10.1631/jzus.2005.B0396

T1 - Effects of medium composition on the production of plasmid DNA vector potentially for human gene therapy
A1 - XU Zhi-nan
A1 - SHEN Wen-he
A1 - CHEN Hao
A1 - CEN Pei-lin
J0 - Journal of Zhejiang University Science B
VL - 6
IS - 5
SP - 396
EP - 400
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Y1 - 2005
PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.2005.B0396

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.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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