2007•Zhongguo shengwuzhipinxue zazhiRequires access
Development of A Procedure for Preparation of Plasmid DNA for Gene Therapy
Ningyi Jin
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Abstract
Objective To explore a procedure for preparation of plasmid DNA for gene therapy.Methods Remove macromolecular RNA from cultured recombinant E.coli with plasmid pIRVP3HNIL18 by calcium chloride precipitation,and other impurities such chromosomal DNA,micromolecular RNA and protein by Q-Sepharose and SOURCE ion exchange chromatography.Results The yield,purity,relative recovery and protein content of final product of plasmid pIRVP3HNIL18 were 1.638 mg plasmid DNA/g bacteria,1.839,12.55% and 0.002 8 mg/ml respectively.No RNA or chromosomal DNA was detected.Conclusion The developed procedure was effective for removal of impurities and suitable for preparation of therapeutic-grade plasmid DNA.