Strain preserving agent and method for anaerobic freezing preservation of genetically engineered bacteria with the same

Abstract

Belonging to the technical field of strain preservation, the invention in particular relates to a strain preserving agent and a method for anaerobic freezing preservation of genetically engineered bacteria with the same. The strain preserving agent comprises the following components by mass: 50-80 parts of a cryoprotectant, 2-5 parts of an antibiotic selective agent and 5-8 parts of a dispersant.The method for anaerobic freezing preservation of genetically engineered bacteria with the preserving agent includes: putting the strain preserving agent into an anaerobic tube, introducing nitrogen and discharging oxygen, then conducting high temperature sterilization; culturing genetically engineered bacteria to a logarithmic growth period, taking fresh bacterial liquid for isovolumetric mixingwith an aseptic anaerobic strain preserving agent, and introducing nitrogen and discharging oxygen in the process, then performing freezing preservation at -20DEG C or -80DEG C. The anaerobic freezingpreservation method can achieve a strict anaerobic condition, reduces the basic metabolic level of strain to the uttermost, can achieve a high resurrection rate, solves the problem of easy lose of genetically engineered bacteria plasmids, and can ensure that the rejuvenated genetically engineered bacteria efficiently maintain the corresponding functions of exogenous DNA fragments.

Description

technical field [0001] The invention belongs to the technical field of strain preservation, and in particular relates to a strain preservation agent and a method for using the preservation agent to anaerobically freeze and preserve genetically engineered bacteria. Background technique [0002] The preservation of strains is essential in microbial research and practical application. Because microorganisms are prone to variation, during the preservation process, the metabolism of microorganisms must be kept in the least active or relatively static state in order to be used for a long time. It does not mutate but is able to maintain viability. Low temperature, dryness and isolation of air are important factors that reduce the metabolic capacity of microorganisms. Although there are many methods for preserving the strains, they are all designed according to these three factors. [0003] Low-temperature cryopreservation is one of the most direct, simplest and most commonly used ...

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problem that genetically engineered bacteria are prone to lose exogenous plasmid replicons in the process of cryopreservation and strain rejuvenation, the present invention provides a strain preservative and a method for using the preservative to anaerobically freeze preserve genetically engineered bacteria