Automatic inducing culture medium for expressing recombinant protein of escherichia coli

A technology for inducing culture medium and recombinant protein, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of unsuitable culture medium, achieve uniformity and consistency, reduce environmental pollution, and increase yield Improved effect

Inactive Publication Date: 2013-09-04
VIVA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional media for recombinant protein production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The frozen pET-ASD glycerol bacteria were selected and inoculated in 100 ml LB medium containing ampicillin at a ratio of 1:1000 and cultured overnight at 37°C. The next day, the bacterial solution was inoculated into 1 liter of the self-inducing medium of the present invention containing ampicillin at a ratio of 1:1000, and 1 liter of the medium contained: tryptone 5g, yeast extract 5g, sodium succinate hexahydrate 0.1 g, sodium citrate dihydrate 0.1g, glycerin 5g, glucose 0.1g, lactose 1g, disodium hydrogen phosphate 2g, potassium dihydrogen phosphate 2g, ammonium chloride 1g, sodium sulfate 0.2g, magnesium sulfate heptahydrate 0.1g, hexahydrate Ferric Chloride 0.1g, Calcium Chloride 1mg, Manganese Chloride Tetrahydrate 0.01mg, Zinc Sulfate Heptahydrate 0.01mg, Cobalt Chloride Hexahydrate 0.01mg, Copper Chloride Dihydrate 0.01mg, Nickel Sulfate Hexahydrate 0.01mg , 0.01 mg of sodium molybdate pentahydrate, 0.04 mg of sodium selenate pentahydrate, 0.01 mg of boric acid...

Embodiment 2

[0020] The frozen pET-APP glycerol bacteria were selected and inoculated in 100 ml of LB medium containing ampicillin and kanamycin at a ratio of 1:1000 and cultured overnight at 37°C. The next day, the bacterial solution was inoculated into 1 liter of the self-inducing medium of the present invention containing ampicillin and kanamycin at a ratio of 1:1000, and 1 liter of the medium contained: 10 g of tryptone, 5 g of yeast extract, six Sodium succinate 8g, sodium citrate dihydrate 1g, glycerin 15g, glucose 1g, lactose 4g, disodium hydrogen phosphate 5g, potassium dihydrogen phosphate 5g, ammonium chloride 4g, sodium sulfate 0.9g, magnesium sulfate heptahydrate 0.1g , ferric chloride hexahydrate 0.4g, calcium chloride 3 mg, manganese chloride tetrahydrate 3 mg, zinc sulfate heptahydrate 5 mg, cobalt chloride hexahydrate 1 mg, copper chloride dihydrate 0.5 mg, nickel sulfate hexahydrate 0.5 mg, 1 mg of sodium molybdate pentahydrate, 0.1 mg of sodium selenate pentahydrate, 0.01...

Embodiment 3

[0022] The frozen pET-BRD glycerol bacteria were selected and inoculated in 100 ml of LB medium containing ampicillin and chloramphenicol at a ratio of 1:1000 and cultured overnight at 37°C. The next day, the bacterial solution was inoculated into 1 liter of the self-inducing medium of the present invention containing ampicillin and chloramphenicol at a ratio of 1:1000, and 1 liter of the medium contained: tryptone 6g, yeast extract 4g, hexahydrate Sodium succinate 3g, sodium citrate dihydrate 0.5g, glycerin 10g, glucose 0.5g, lactose 2g, disodium hydrogen phosphate 2.5g, potassium dihydrogen phosphate 2.5g, ammonium chloride 3g, sodium sulfate 0.6g, sulfuric acid heptahydrate Magnesium 0.5g, ferric chloride hexahydrate 0.2g, calcium chloride 2mg, manganese chloride tetrahydrate 1mg, zinc sulfate heptahydrate 2mg, cobalt chloride hexahydrate 0.5mg, copper chloride dihydrate 0.1mg, hexahydrate Nickel sulfate water 0.1 mg, sodium molybdate pentahydrate 0.1 mg, sodium selenate pe...

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Abstract

The invention relates to an automatic inducing culture medium for expressing recombinant protein of escherichia coli. Each liter of the culture medium comprises components in parts by weight as follows: 5-10g of tryptone, 3-5g of a yeast extract, 0.1-8 g of sodium succinate hexahydrate, 0.1-1g of sodium citrate dehydrate, 5-15g of glycerinum, 0.1-1g of glucose, 1-4g of lactose, 2-5g of disodium hydrogen phosphate, 2-5g of monopotassium phosphate, 1-4g of ammonium chloride, 0.2-0.9g of sodium sulfate, 0.1-1g of magnesium sulfate heptahydrate, 0.1-0.4g of ferric chloride hexahydrate, 1-3mg of calcium chloride, 0.01-3mg of tetrahydrate manganese chloride, 0.01-5mg of zinc sulfate heptahydrate, 0.01-1mg of cobalt chloride hexahydrate, 0.01-0.5mg of copper chloride dehydrate, 0.01-0.5mg of nickel sulfate hexahydrate, 0.01-1mg of sodium molybdate pentahydrate, 0.04-0.1mg of sodium selenate pentahydrate, 0.001-0.01mg of boric acid, 1-100g of n-hexane and 0.1-10g of calcium peroxide. The automatic inducing culture medium is used for expressing foreign protein, the expression amount of the foreign recombinant protein of the automatic inducing culture medium is increased when compared with that of foreign recombinant protein of an LB (Luria-Bertani) ordinary culture medium, the production cost is saved, environmental pollution is reduced, and the yield can be increased by more than ten times.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a culture medium, in particular to an automatic induction culture medium for expressing recombinant proteins in Escherichia coli. Background technique [0002] Bacteria have been used to produce antibiotics since the mid-20th century, but these products have tended to be limited to compounds the bacteria naturally synthesize. In the past 20 years, with the popularization and promotion of gene cloning technology, people have used bacteria to produce a large number of recombinant proteins. [0003] LB medium is generally used to cultivate Escherichia coli, but LB medium has a series of disadvantages when expressing recombinant proteins. One: One of the main components of LB medium is tryptone, which is inevitably mixed with a small amount of lactose, resulting in non-target bacteria. Express. Especially when expressing toxic proteins, bacteria are easy to die, resulting in the failure ...

Claims

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Application Information

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IPC IPC(8): C12P21/00C12R1/19
Inventor 田为宇薛飞樊盼盼彭金河尚建强谭恩旺赵岩龙方琼
Owner VIVA BIOTECH
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