Culture medium and culture method for efficiently expressing plasmid DNA through Escherichia coli engineering bacteria

Abstract

The present invention provides a culture medium and a culture method for efficiently expressing plasmid DNA through Escherichia coli engineering bacteria. The culture medium comprises a base culture medium and a material supplementing culture medium, wherein the base culture medium comprises: 7.75 ml/L of glycerol, 3.0 g/L of ammonium sulfate, 6.0 g/L of dipotassium hydrogen phosphate, 3.0 g/L of potassium dihydrogen phosphate, 1.7 g/L of citric acid, 1.2 g/L of magnesium sulfate, 30 mg/L of ferric chloride, 5.00 g/L of a yeast extract, 1.00 mg/L of D-biotin, and 9.70 Ml/L of a trace element solution, and the material supplementing culture medium comprises 10 ml/L of glycerol, 3.0 g/L of ammonium sulfate, 6.0 g/L of dipotassium hydrogen phosphate, 3.0 g/L of potassium dihydrogen phosphate, 1.7 g/L of citric acid, 1.2 g/L of magnesium sulfate, 30 mg/L of ferric chloride, 20.00 g/L of a yeast extract, and 1.00 mg/L of D-biotin. The culture method uses a fed-batch mode, and can significantly improve the total content of the fermented plasmid on the basis of the maintaining of the high plasmid content per unit bacteria.

Description

technical field [0001] The invention belongs to the field of biology, and relates to a culture medium and a culture method for high-efficiency expression of plasmid DNA by Escherichia coli engineering bacteria. Background technique [0002] With the development of recombinant DNA technology and genome sequencing in recent years, significant progress has been made in the fields of gene therapy and DNA vaccine research. Plasmid DNA carrying exogenous genes is the main drug carrier used in gene therapy and DNA vaccine research and development. At present, the clinical dosage of therapeutic plasmid DNA per patient has reached the mg level, so the large-scale preparation of plasmid DNA requires industrial-scale E. coli engineering bacteria fermentation technology to prepare. In terms of fermentation technology, the early plasmid fermentation medium was derived from the medium of recombinant protein genetically engineered bacteria. However, unlike the engineering bacteria expres...

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Problems solved by technology

[0005] Invention patent application 2009102137258 A medium for increasing the specific content of plasmid DNA in Escherichia coli engineering bacteria can significantly increase the total plasmid co

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