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Large-scale plasmid preparation process

A technology for large-scale preparation of plasmids, applied in the field of bioengineering, can solve problems such as difficulties in large-scale preparation of plasmids, and achieve the effects of simple methods, convenient operation, and reduced production costs

Inactive Publication Date: 2015-08-12
SHANDONG BINZHOU ANIMAL SCI & VETERINARY MEDICINE ACADEMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] One of the purposes of the present invention is to provide a convenient and simple large-scale preparation process for plasmids in order to solve the difficult problem of large-scale preparation of plasmids

Method used

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Embodiment Construction

[0028] The present invention will be described in detail below in conjunction with specific embodiments. It should be noted that the technical features or combinations of technical features described in the following embodiments should not be regarded as isolated, and they can be combined with each other to achieve better technical effects.

[0029] The invention provides a large-scale preparation process of a plasmid, comprising the following steps:

[0030] Step 1: Preparation of the seed batch

[0031] 1) Separate and purify the transformed K12DH-5α freeze-dried strain by streaking LB agar plate, and identify the recombinant plasmid by restriction enzyme digestion and PCR. Take typical colonies and streak LB agar plates, culture overnight at 37°C, wash with 0.01mol / L, pH7.2 PBS solution to LB liquid medium, fill in 5mL sterilized vials, add 10% glycerol and mix well, as a Grade seeds, stored at -70°C. Inoculate first-grade seeds into LB medium containing 30 μg / mL kanamyc...

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Abstract

The invention discloses a large-scale plasmid preparation process which comprises the following steps: preparing production seeds, performing high-density fermentation, thalli harvesting, removing RNA and extracting and purifying the plasmids. The large-scale plasmid preparation process disclosed by the invention is simple in method and convenient to operate, lots of plasmids can be produced, and the production cost is reduced.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a large-scale preparation process of plasmids. Background technique [0002] DNA vaccine is a new type of vaccine developed in the 1990s, and it is the third generation of vaccine after attenuated vaccine and genetic engineering vaccine. DNA vaccine (DNAvaccine) is composed of elements such as plasmid DNA (from bacteria) inserted with one or more foreign genes and eukaryotic promoter regulatory genes. Under the control of gene units such as tailing signals and related enhancers, related antigenic proteins can be expressed in various cells of mammals. The recombinant plasmid with the gene encoding the foreign antigen is directly introduced into human or animal cells by a certain method, and the antigen protein is synthesized in the living cells of the immunized body through the transcription system of the host cell, thereby inducing the body to produce an immune response...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/10
Inventor 郭广君吕素芳沈志强吴信明付强王艳
Owner SHANDONG BINZHOU ANIMAL SCI & VETERINARY MEDICINE ACADEMY
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