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Culture medium for Escherichia coli capable of efficiently expressing plasmid DNA and preparation method of culture medium

A high-efficiency expression and Escherichia coli technology, applied in the biological field, can solve the problems of low plasmid DNA content and unsatisfactory results, and achieve the effect of increasing the amount of bacteria per unit and improving the preparation

Inactive Publication Date: 2019-02-22
安徽欣伯玉生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although the existing medium used for high-density expression of plasmid DNA engineering bacteria can achieve a higher bacterial density and total plasmid content, the relative content of plasmid DNA is generally low. Using the existing medium formula, although the plasmid DNA content has improved to some extent, but the results are still unsatisfactory

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  • Culture medium for Escherichia coli capable of efficiently expressing plasmid DNA and preparation method of culture medium

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Experimental program
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Effect test

Embodiment 1

[0025] A medium for efficiently expressing plasmid DNA in Escherichia coli, consisting of the following parts by weight:

[0026] 2 parts beef extract, 10 parts peptone, 2 parts sodium chloride, 3 parts modified lanthanum carbonate, 1 part activated carbon, 0.3 part tartaric acid, 0.1 part citric acid, 12 parts glycerin, 800 parts deionized water.

[0027] Further, the preparation method of the modified lanthanum carbonate includes the following steps:

[0028] a. Put the lanthanum carbonate into a 2% hydrochloric acid solution for soaking, keep the temperature in the hydrochloric acid solution at 60°C during soaking, and filter out after soaking for 10 minutes;

[0029] b. Put the lanthanum carbonate soaked in the acid solution in step a into a sodium hydroxide solution with a mass fraction of 4% for soaking, keep the temperature in the sodium hydroxide solution at 90°C during soaking, and filter out after soaking for 17 minutes. ;

[0030] c. Put the lanthanum carbonate soaked in sod...

Embodiment 2

[0039] A medium for efficiently expressing plasmid DNA in Escherichia coli, consisting of the following parts by weight:

[0040] 3 parts of beef extract, 11 parts of peptone, 2.5 parts of sodium chloride, 3.5 parts of modified lanthanum carbonate, 1.5 parts of activated carbon, 0.35 parts of tartaric acid, 0.15 parts of citric acid, 13 parts of glycerin, and 850 parts of deionized water.

[0041] Further, the preparation method of the modified lanthanum carbonate includes the following steps:

[0042] a. Put lanthanum carbonate into a hydrochloric acid solution with a mass fraction of 3% for soaking, keep the temperature in the hydrochloric acid solution at 65°C during soaking, and filter out for use after soaking for 11 minutes;

[0043] b. Put the lanthanum carbonate soaked in the acid solution in step a into a 4.5% sodium hydroxide solution for soaking, keep the temperature in the sodium hydroxide solution at 95°C during soaking, and filter out after soaking for 18 minutes. ;

[00...

Embodiment 3

[0053] A medium for efficiently expressing plasmid DNA in Escherichia coli, consisting of the following parts by weight:

[0054] 4 parts beef extract, 12 parts peptone, 3 parts sodium chloride, 4 parts modified lanthanum carbonate, 2 parts activated carbon, 0.4 parts tartaric acid, 0.2 parts citric acid, 14 parts glycerin, 900 parts deionized water.

[0055] Further, the preparation method of the modified lanthanum carbonate includes the following steps:

[0056] a. Put lanthanum carbonate into a hydrochloric acid solution with a mass fraction of 4% for soaking, keep the temperature in the hydrochloric acid solution at 70°C during soaking, and filter out after soaking for 12 minutes;

[0057] b. Put the lanthanum carbonate after soaking in the acid solution in step a into a sodium hydroxide solution with a mass fraction of 5% for soaking. Keep the temperature in the sodium hydroxide solution at 100°C during soaking, and filter out after soaking for 19 minutes. ;

[0058] c. Put the la...

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Abstract

The invention discloses a culture medium for Escherichia coli capable of efficiently expressing plasmid DNA and a preparation method of the culture medium. The culture medium is composed of the following components in parts by weight: beef extract, peptone, sodium chloride, modified lanthanum carbonate, activated carbon, tartaric acid, citric acid, glycerin and deionized water. According to the finally prepared culture medium disclosed by the invention, preparation of the conventional LB culture medium is effectively improved, the unit microbial quantity in a unit culture medium is improved, and the cultured Escherichia coli is capable of efficiently expressing the plasmid DNA, lays a good foundation for development of genetic engineering, and has excellent market popularization value.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a culture medium for efficiently expressing plasmid DNA in Escherichia coli and a preparation method thereof. Background technique [0002] Plasmids have the ability to replicate autonomously, allowing them to maintain a constant copy number in progeny cells and express the genetic information they carry. Bacterial plasmid is a commonly used vector in DNA recombination technology. A vector is a tool for sending a useful foreign gene into the recipient cell for proliferation and expression through genetic engineering. Recombination of a certain target gene fragment into a plasmid to form a recombinant gene or recombinant body. Then the recombinant is transformed into the recipient cell (such as Escherichia coli) by microbiological transformation technology, so that the target gene in the recombinant can be reproduced or expressed in the recipient bacteria, thereby changing the...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12R1/19
CPCC12N1/20
Inventor 赵建阳王世凯韦小艳
Owner 安徽欣伯玉生物科技有限公司