Method of continuous large quantity extracting plasmid

Abstract

This invention discloses a preparation of continuously and greatly extracting plasmid. This preparation is that using reagent of schizolysis bacterium thallus to split bacterium thallus, obtains thallus lysate; then adding reagent used for precipitating protein and genome DNA into stated thallus lysate, obtains the mixed liquor which contains protein sediment and genome DNA sediment; then this mixed liquor is filtered, protein sediment and genome DNA sediment which are in the stated mixed liquor are removed, filter liquor is collected, plasmid is obtained.

Description

Technical field [0001] The invention relates to a method for continuously extracting a large amount of plasmids. Background technique [0002] Nucleic acid vaccines and gene therapy have made rapid progress in recent years (Liljeqvist, S. and Stahl, S, 1999. Production of recombinant subunit vaccines: protein immunogens, live delivery and nucleic acid vaccines. J. Biotech. 73, 1-33; Friedman , T, 1997. The road toward human gene therapy-a 25-year perspective. Ann. Med. 29, 575-577). However, the efficiency of plasmid-based delivery vectors to transfect cells is very low—only 0.1% of the plasmids can reach the nucleus and be expressed (Crystal, R.G., 1995. The gene as the drug. Nat. Med. 1, 15-17). With the clinical application of nucleic acid vaccines and gene therapy, more and more plasmids are needed in production and scientific research (Guilherme NMFerreira, Gabriel A. Monteiro, Duarte MFPrazeres and Joaquim MSCabral, 2000. Downstream processing of plasmid DNA for genetherapy...

AI Technical Summary

Problems solved by technology

Due to the higher requirements for temperature control, the boiling method is less stable in the plasmid preparation of the laboratory (Wang, B., Merva, M., Williams, W., Weiner, D., 1995.Large-scale preparation of plasmid DNA by microwave analysis.Biotechniques 18, 554-555), not to mention industrial production

In 2005, Zhu Kaichun and others invented and optimized a continuous thermal cracking method (Kaichun Zhu, Huali Jin, Yijie Ma, Zhihui Ren, Chong Xiao, Zhonghuai He, Fuchun Zhang, Qinghong Zhu and Bin Wang, 2005.Acontinuous thermal lysis procedure for the large-scale preparation of plasmaDNA. Journal of Biotechnology. 118, 257-264), this method improves the traditional boiling method, the whole process is continuous, can handle a large number of bacteria and save costs, However, the post-extraction process in this method is still difficult

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