CN114426943A - Escherichia coli expression culture medium for expression of novel coronavirus N protein - Google Patents
Escherichia coli expression culture medium for expression of novel coronavirus N protein Download PDFInfo
- Publication number
- CN114426943A CN114426943A CN202210206163.XA CN202210206163A CN114426943A CN 114426943 A CN114426943 A CN 114426943A CN 202210206163 A CN202210206163 A CN 202210206163A CN 114426943 A CN114426943 A CN 114426943A
- Authority
- CN
- China
- Prior art keywords
- culture medium
- protein
- escherichia coli
- expression
- novel coronavirus
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/20011—Coronaviridae
- C12N2770/20022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Virology (AREA)
- Biophysics (AREA)
- Tropical Medicine & Parasitology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- General Engineering & Computer Science (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention belongs to the technical field of biology, and particularly relates to an escherichia coli expression culture medium for expressing a novel coronavirus N protein. The invention provides a culture medium comprising peptone and yeast extract, but not NaCl, but adding an A reagent which can be C4H11NO3HCl. The amount of hybrid protein in the expressed protein of the escherichia coli BL21 containing the novel coronavirus N protein gene cultured by using the culture medium prepared by the invention is greatly reduced, and the difficulty of later-stage purification is reduced.
Description
The application is a divisional application with the patent application number of '202110267795.2', the application date of '3.11.2021', the invention name of 'an escherichia coli expression culture medium for expressing a novel coronavirus N protein'.
Technical Field
The invention belongs to the technical field of biology, and particularly relates to an escherichia coli expression culture medium for expressing a novel coronavirus N protein.
Background
The Escherichia coli is one of the main hosts for exogenous gene expression, has wide application, and has the advantages of clear genetic background, simple technical operation, simple culture condition, strong pollution resistance, large-scale fermentation economy and the like. 2019-nCoV has strong infectivity. The protein is divided into: spike glycoprotein (S protein), envelope glycoprotein (E protein), membrane glycoprotein (M protein), and nucleocapsid protein (N protein). To obtain the novel coronavirus N protein, a plasmid with a foreign gene capable of expressing the N protein is transformed into escherichia coli for induction expression and purification. Usually, the fermentation medium of escherichia coli is prepared as an LB medium, but after escherichia coli with an N protein foreign gene is induced and expressed in the LB medium, the produced protein has a large amount of hybrid protein besides the N protein, wherein part of the hybrid protein cannot be removed through various purification modes.
Disclosure of Invention
Aiming at the defects generally existing in the prior art, the invention provides an escherichia coli expression culture medium for expressing a novel coronavirus N protein. The amount of hybrid protein in the expressed protein of the escherichia coli BL21 containing the novel coronavirus N protein gene cultured by using the culture medium prepared by the invention is greatly reduced, and the difficulty of later-stage purification is reduced.
In order to achieve the purpose, the invention adopts the technical scheme that:
an escherichia coli expression culture medium for expression of a novel coronavirus N protein comprises peptone, yeast extract and an A reagent.
Preferably, the addition amount of the peptone is 10g, and the addition amount of the yeast extract is 5 g.
Preferably, the reagent A is C4H11NO3·HCl。
Preferably, said C4H11NO3The final HCl mass concentration is 1.576 to 4.728 g/L.
The invention also provides a preparation method of the culture medium, which comprises the following steps: weighing the components in the culture medium by using a balance, dissolving the components in 900mL of purified water, uniformly mixing, adjusting the pH value to 8.5 by using 1mol/L NaOH solution, fixing the volume to 1L by using the purified water, and autoclaving at 121 ℃ for 20min to obtain the culture medium.
The culture medium provided by the invention is a culture medium which is obtained by improving an LB liquid culture medium through a large number of experiments, and removes NaCl originally contained in the LB liquid culture medium, so that the amount of the hybrid protein in the protein expressed by escherichia coli is reduced, and the subsequent protein purification is facilitated.
Compared with the prior art, the culture medium provided by the invention has the following advantages: when the culture medium of the formula is used for culturing the escherichia coli BL21 containing the novel coronavirus N protein gene, the amount of hybrid protein in the expressed protein is reduced, and the content of a hybrid band near 23kD is obviously reduced. The influence of non-specific protein on subsequent purification is reduced, and the difficulty of later purification is reduced.
Drawings
FIG. 1 is a diagram showing the results of the example of the E.coli expression medium of the present invention.
Detailed Description
The present invention is further explained with reference to the following specific examples, but it should be noted that the following examples are only illustrative of the present invention and should not be construed as limiting the present invention, and all technical solutions similar or equivalent to the present invention are within the scope of the present invention. The method and the device are operated according to the conventional technical method and the content of the instrument instruction, wherein the specific technology or condition is not indicated in the embodiment; the reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
The preparation process of each reagent involved in the invention is as follows:
preparation of 50mg/mL Kan (kanamycin) solution: weighing 0.5g kanamycin by using an analytical balance, dissolving by using ultrapure water, diluting to a constant volume of 10mL, filtering and sterilizing by using a 0.22-micron filter membrane, subpackaging into 1.5mL centrifuge tubes, marking antibiotic names, stock solution concentration and preparation date, and storing in a lightproof container at-20 ℃. The working solution concentration was 50. mu.g/mL.
② 1mol/L IPTG (isopropyl-beta-D-thiogalactoside) solution preparation: weighing 2.3830g of IPTG, ultra-pure water for dissolving and fixing the volume to 10mL by using an analytical balance, filtering and sterilizing by using a 0.22-micron filter membrane, subpackaging into 1.5mL centrifuge tubes, marking the name of a solution, the concentration of a stock solution and the preparation date, and storing at-20 ℃.
Preparation of 1L LB solid medium: 10g of peptone, 5g of yeast extract, 10g of sodium chloride and 15g of agar powder, dissolving with purified water and fixing the volume to 1L. Autoclaving at 121 deg.C for 20min, and cooling to 55 deg.C. Adding antibiotic solution (Kan) in a corresponding proportion according to the required resistance concentration, pouring the plate according to the aseptic operation standard, airing, marking the resistance type and preparation date of the plate, sealing by a sealing film, and storing for 2 months in an inverted manner at 4 ℃. Remarking: the solid culture medium added with antibiotics can be stored for half a month at 4 ℃, is only used for culturing a single bacterial colony, and is a selective liquid culture medium in the following bacterial culture.
Preparing 1L of LB liquid culture medium: dissolving 10g of peptone, 5g of yeast extract and 10g of sodium chloride in purified water, diluting to a constant volume of 1L, and autoclaving at 121 deg.C for 20min to obtain the final product.
Preparing 30% (W/V) acrylamide: 290 g of acrylamide and 10g of methylene bisacrylamide are weighed in a beaker, 600 mL of deionized water is added, the mixture is fully stirred and dissolved, the volume is determined to be 1L by the deionized water, and the mixture is stored at 4 ℃ in a dark place. Note that: acrylamide has strong neurotoxicity, can be absorbed through skin, has accumulative effect, and can be used by wearing gloves during preparation.
Sixthly, preparing a separation gel buffer solution (1.5M Tris, pH 8.8): 181.6 g Tris is weighed into a beaker, 800 mL deionized water is added, the mixture is fully stirred and dissolved, hydrochloric acid is added to adjust the PH value to 8.8, deionized water is used for fixing the volume to 1L, and the mixture is stored at room temperature.
Concentrating the gum buffer (1M Tris, pH 6.8): weighing 121.1 g of Tris in a beaker, adding 800 mL of deionized water, fully stirring and dissolving, adding hydrochloric acid to adjust the pH value to 6.8, using deionized water to fix the volume to 1L, and storing at room temperature.
(viii) 10% (W/V) SDS: 10g SDS was weighed into a beaker, 100 mL deionized water was added, and dissolved with stirring.
Ninthly 10% (W/V) ammonium persulfate: weighing 1g of ammonium persulfate into a centrifuge tube, adding 10mL of deionized water, stirring for dissolving, and storing at 4 ℃ or-20 ℃ (4 ℃ can be stored for 2 weeks and-20 ℃ can be stored for 1 month).
shortSDS-PAGE electrophoresis Buffer (10 × Tris-Glycine Buffer): 30.3g of Tris, 144 g of Glycine and 10g of SDS are weighed and placed in a beaker, about 800 mL of deionized water is added, stirring and dissolving are carried out, deionized water is added to the solution, the volume is fixed to 1L, and then the solution is stored at room temperature.
⑪ SDS-PAGE Loading buffer (5X): 0.5g SDS and 25mg bromophenol blue are weighed and placed in a plastic centrifuge tube, 1.25 mL Tris-HCI (1M, PH6.8) and 2.5 mL glycerol are absorbed in the centrifuge tube, deionized water is added for dissolution, the volume is determined to be 5mL, and the small portions are packaged at room temperature. 5% (volume ratio) of beta-mercaptoethanol (capable of being stored at room temperature for 1 month) is added before use.
⑫ Coomassie brilliant blue R-250 staining solution: weighing 1g of Coomassie brilliant blue R-250, placing the Coomassie brilliant blue R-250 in a beaker, weighing 250 mL of isopropanol, adding the isopropanol into the beaker, stirring for dissolving, adding 100 mL of glacial acetic acid, stirring uniformly, adding 650 mL of deionized water, stirring uniformly, removing particulate matters by using filter paper, and storing at room temperature.
⑬ Coomassie brilliant blue staining decolorized solution: 100 mL of acetic acid and 50mL of ethanol were weighed into a beaker, 850 mL of deionized water was added to the beaker, and the mixture was mixed thoroughly.
Example 1 E.coli expression Medium for the expression of the N protein of a novel coronavirus
The culture medium comprises 10g of peptone, 5g of yeast extract and 1.576g C4H11NO3·HCl。
The preparation process of the culture medium comprises the following steps: dissolving the above components with 900mL of purified water, adjusting pH to 8.5 with 1mol/L sodium hydroxide solution, diluting to 1L with purified water, and sterilizing at 121 deg.C under 0.12MPa for 20 min.
Example 2 E.coli expression Medium for the expression of the N protein of a novel coronavirus
The culture medium comprises 10g of peptone, 5g of yeast extract and 3.152g C4H11NO3·HCl；
The preparation of the medium was carried out analogously to example 1.
Example 3 E.coli expression Medium for the expression of the N protein of a novel coronavirus
The culture medium comprises 10g of peptone, 5g of yeast extract and 4.728g C4H11NO3·HCl；
The preparation of the medium was carried out analogously to example 1.
Example 4 E.coli expression Medium for the expression of the N protein of a novel coronavirus
The medium included 10g peptone, 5g yeast extract, 1.45 g disodium hydrogen phosphate, 0.13g potassium dihydrogen phosphate.
The preparation process of the culture medium comprises the following steps: adding the components into 900mL of purified water for dissolving, then continuing to add the purified water to a constant volume of 1L, and placing at 121 ℃ for autoclaving for 20min to obtain the composition.
Example 5 E.coli expression Medium for the expression of the N protein of a novel coronavirus
The medium comprises 10g of peptone, 5g of yeast extract, 2.9g of disodium hydrogen phosphate, and 0.26g of potassium dihydrogen phosphate;
the preparation of the medium was carried out analogously to example 4.
Example 6 E.coli expression Medium for the expression of the N protein of a novel coronavirus
The medium comprises 10g of peptone, 5g of yeast extract, 5.8g of disodium hydrogen phosphate, and 0.52g of potassium dihydrogen phosphate;
the preparation of the medium was carried out analogously to example 4.
Experimental example Strain culture and verification Process thereof
1. Test samples: example 1-6 groups of formulated medium.
2. The test process comprises the following steps: the specific test process is as follows:
before a specific test, a plasmid with a foreign gene capable of expressing N protein is transformed into escherichia coli, and the specific process comprises the following steps: the registered new crown N protein sequence is searched from GenBank, and the sequence is sent to a synthetic plasmid of Nikkita. The plasmid vector was PET-28A (+) at a concentration of 500 ng/. mu.L. mu.L of the plasmid was transformed into competent E.coli BL21 by heat shock, incubated on ice for 5 minutes, then added with 300. mu.L of LB without resistance to allow resuscitation for 10min (shaker 37 ℃, 220 rpm), spread on LB plate containing 50. mu.g/ml Kan +, and incubated overnight at 37 ℃ in an incubator. The next day, 1 positive clone was picked into 5mL of fresh LB liquid medium containing 50. mu.g/mL Kan +, incubated for 8 hours at 37 ℃ and 220rpm in a shaker, 600. mu.L of bacterial liquid was taken, 300. mu.l of 70% sterilized glycerol was added, and the mixture was mixed uniformly to obtain the strain used in the present invention, which was frozen in a freezer at-80 ℃.
(1) Taking the 2019-nCoV-2-N strain out of a refrigerator at the temperature of-80 ℃ and placing the strain on an ice box;
(2) scribing a flat plate: taking part of strains by using a sterilized inoculating needle, streaking the strains on a solid kana resistant plate, and inversely placing the streaked strains in an LB solid incubator at 37 ℃ for culturing for 14 h;
(3) after the culture is finished, picking single colonies into 5mL respectively, placing the single colonies into LB liquid culture medium (control culture medium) with the final concentration of 50 mug/mL Kana and 6 groups of culture mediums of examples, and placing the culture mediums into constant temperature shake culture at 37 ℃ for 14 h;
(4) inoculating each bacterial liquid cultured in the step (3) into a fresh LB liquid culture medium (contrast culture medium) with the final concentration of 50 mu g/mL Kana resistance and the volume of 50mL and 6 groups of culture media of examples in sequence according to the proportion of 2% (V/V), and carrying out constant-temperature shaking culture at 37 ℃ and 260rpm (measuring the absorbance value of the bacterial liquid OD600 by Nano 300);
(5) when the OD600 reaches 0.6-0.8, adding 50 mu L of IPTG with the concentration of 1mol/L, and inducing for 10h at 37 ℃ and 220 rpm;
(6) placing the induced product at 25 ℃, centrifuging at 9000rpm for 3min, and placing the recovered thallus in a sterile 50mL centrifuge tube;
(7) adding 3mL of 20mM Tris-HCl (pH8.5) into a centrifuge tube to wash the thalli once, carrying out vortex oscillation, centrifuging for 3min at 25 ℃ and 9000rpm, collecting the thalli, and marking the strain name and the experimental date;
(8) washing 1mL of the collected induced thallus once more with 20mM Tris-HCl (pH8.5), centrifuging at 25 deg.C and 12000rpm for 1min, and collecting thallus precipitate again;
(9) adding 100 μ L of 20mM Tris-HCl (pH8.5) into the bacterial pellet to resuspend the bacterial pellet;
(10) mixing SDS-PAGE sample buffer (5 x) and the bacterial suspension at a ratio of 1:4, heating on a dry heating instrument at 100 ℃ for 10min to prepare a sample to be detected;
(11) preparing SDS-PAGE protein electrophoresis gel: colloids with corresponding concentrations were prepared according to the ratios described in tables 1 and 2 below.
TABLE 15% PAGE gel concentrate formulation
TABLE 210% PAGE gel formulation
(12) Sequentially adding 20 mu L of prepared samples into the glue holes, and making related experimental records;
(13) sample concentration process, setting electrophoresis conditions: 100V, 80mA (10% SDS-PAGE gel, the SDS-PAGE coagulates two layers, the upper layer is concentrated glue, the lower layer is separation glue, when in use, after the separation glue of the lower layer coagulates, the layer of concentrated glue is added for coagulation);
(14) when the sample indicator tape is electrophoresed to the interface of the concentrated gel and the separation gel, the electrophoresis conditions are adjusted to: 130V, 100 mA;
(15) when the indicator tape is electrophoresed to be about 1cm away from the bottom of the rubber plate, the electrophoresis is stopped, and the power supply is turned off;
(16) dyeing glue: and taking out the rubber plate, sorting out the colloid by using a colloid extractor, and placing the colloid in the dyeing liquid to ensure that the dyeing liquid is sufficient. Placing the box containing the staining solution in a rotary horizontal shaking table, and oscillating at room temperature and 80rpm for 40 min;
(17) and (3) decoloring: the colloid in the dyeing liquid is taken out and is slowly washed twice by ultrapure water, the colloid is prevented from being washed out, then the colloid is transferred into a box containing enough decolorizing liquid, the box is placed in a rotary horizontal shaking table, the box is oscillated for 2 hours at the room temperature of 80rpm, and 1-2 times of decolorizing liquid is replaced during the process so as to be rapidly decolorized.
3. And (3) test results: the specific test results are shown in FIG. 1, the SDS-PAGE gel is 5% of concentrated gel, the separation gel is 10% of separation gel, and the control medium is liquid LB medium. Marker was purchased from Shanghai Yazyme biology Inc.; as can be seen from Table 1, the N protein bands in the bacterial solutions prepared by using the culture mediums of examples 1-6 of the present invention were significantly lower than those in the control culture medium.
Finally, it should be noted that the above description of embodiments is provided to facilitate the understanding and use of the invention by those skilled in the art. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make modifications and alterations without departing from the scope of the present invention.
Claims (2)
1. An escherichia coli expression culture medium for expressing a novel coronavirus N protein is characterized by comprising peptone, yeast extract, a reagent A and water, wherein the addition amount of the peptone is 10g and the addition amount of the yeast extract is 5g in each liter of culture medium; the reagent A is C4H11NO3HCl, said C4H11NO3The final mass concentration of HCl is 1.576-4.728 g/L; the balance being water.
2. A method for preparing the culture medium of claim 1, which comprises the following steps: weighing the components in the culture medium by using a balance, dissolving the components in 900mL of purified water, uniformly mixing, adjusting the pH value to 8.5 by using 1mol/L NaOH solution, fixing the volume to 1L by using the purified water, and autoclaving at 121 ℃ for 20min to obtain the culture medium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210206163.XA CN114426943B (en) | 2021-03-11 | 2021-03-11 | Escherichia coli expression culture medium for expression of novel coronavirus N protein |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210206163.XA CN114426943B (en) | 2021-03-11 | 2021-03-11 | Escherichia coli expression culture medium for expression of novel coronavirus N protein |
| CN202110267795.2A CN113151128B (en) | 2021-03-11 | 2021-03-11 | Escherichia coli expression culture medium for expression of novel coronavirus N protein |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110267795.2A Division CN113151128B (en) | 2021-03-11 | 2021-03-11 | Escherichia coli expression culture medium for expression of novel coronavirus N protein |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114426943A true CN114426943A (en) | 2022-05-03 |
| CN114426943B CN114426943B (en) | 2022-11-11 |
Family
ID=76887013
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210206163.XA Active CN114426943B (en) | 2021-03-11 | 2021-03-11 | Escherichia coli expression culture medium for expression of novel coronavirus N protein |
| CN202110267795.2A Active CN113151128B (en) | 2021-03-11 | 2021-03-11 | Escherichia coli expression culture medium for expression of novel coronavirus N protein |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110267795.2A Active CN113151128B (en) | 2021-03-11 | 2021-03-11 | Escherichia coli expression culture medium for expression of novel coronavirus N protein |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN114426943B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5037756A (en) * | 1986-04-21 | 1991-08-06 | The United States Of America As Represented By The Secretary Of The Army | Recombinant DNA molecules for producing terminal transferase-like polypeptides |
| CN105671113A (en) * | 2016-03-08 | 2016-06-15 | 四川省农业科学院土壤肥料研究所 | Culture medium for promoting Escherichia coli to secrete expression recombinant proteins and preparation method thereof |
| US20200207815A1 (en) * | 2016-12-06 | 2020-07-02 | "Future Analgesics" Limited | Peptide modulator of purinergic receptors |
| CN112280794A (en) * | 2020-11-03 | 2021-01-29 | 安徽环球基因科技有限公司 | Production method of new crown recombinant NP protein in escherichia coli |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101235362B (en) * | 2007-02-01 | 2010-08-11 | 中国农业科学院哈尔滨兽医研究所 | Composite automatic induction culture medium for expressing exogenous protein by prokaryocyte expression system |
| CN103276032A (en) * | 2013-06-09 | 2013-09-04 | 维亚生物科技(上海)有限公司 | Automatic inducing culture medium for expressing recombinant protein of escherichia coli |
-
2021
- 2021-03-11 CN CN202210206163.XA patent/CN114426943B/en active Active
- 2021-03-11 CN CN202110267795.2A patent/CN113151128B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5037756A (en) * | 1986-04-21 | 1991-08-06 | The United States Of America As Represented By The Secretary Of The Army | Recombinant DNA molecules for producing terminal transferase-like polypeptides |
| CN105671113A (en) * | 2016-03-08 | 2016-06-15 | 四川省农业科学院土壤肥料研究所 | Culture medium for promoting Escherichia coli to secrete expression recombinant proteins and preparation method thereof |
| US20200207815A1 (en) * | 2016-12-06 | 2020-07-02 | "Future Analgesics" Limited | Peptide modulator of purinergic receptors |
| CN112280794A (en) * | 2020-11-03 | 2021-01-29 | 安徽环球基因科技有限公司 | Production method of new crown recombinant NP protein in escherichia coli |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113151128A (en) | 2021-07-23 |
| CN114426943B (en) | 2022-11-11 |
| CN113151128B (en) | 2022-04-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Angus | Extraction, purification, and properties of Bacillus sotto toxin | |
| AU594062B2 (en) | Bacterial enzymes | |
| JPS58126789A (en) | Method for developing genetic character | |
| DE3686579T2 (en) | PROKARYOTIC EXPRESSION SYSTEM. | |
| WO1987005932A1 (en) | Biological containment | |
| KR20080018960A (en) | A method of secretion expression of lysostaphin in escherichia coli at high level | |
| CN113151128B (en) | Escherichia coli expression culture medium for expression of novel coronavirus N protein | |
| CN107034226A (en) | A kind of soluble recombinant protein and its expression and purification method and purposes | |
| BR9810650B1 (en) | vector for heterologous protein expression and methods for extracting recombinant protein and for purifying isolated recombinant insulin. | |
| CN113045670B (en) | Soluble chicken alpha interferon fusion protein and production method and application thereof | |
| CN110687035A (en) | Annexin V-Light650 apoptosis detection kit | |
| CN114736273B (en) | Phage alpha3 cleavage protein E and application thereof | |
| JP4372421B2 (en) | Methods and compositions for extracting proteins from cells | |
| CN116284317A (en) | Adiponectin ADP renaturation method with high stability and high reactivity | |
| CN110846326A (en) | Raccoon parvovirus VP2 gene, expression vector, recombinant strain, method for preparing VP2 protein and assembly method | |
| Krügel et al. | Transfection of protoplasts from Streptomyces lividans 66 with actinophage SH10 DNA | |
| CN115873077A (en) | Reagent for detecting bovine nodular skin disease virus antibody and polypeptide used by reagent | |
| JP3142348B2 (en) | Recombinant plasmid having nitrile-degrading enzyme gene, transformed microorganism, and method for producing amide and acid using the transformed microorganism | |
| CN115637247A (en) | Efficient biological oxidation desulfurization halomonas engineering strain and whole-cell catalytic desulfurization method thereof | |
| CN105950629A (en) | Prokaryotic expression vector culture method and method for preparing antiserum through prokaryotic expression vector | |
| CN111057155A (en) | O-type FMDV VP1 protein-ferritin fusion protein, protein cage nanoparticle and preparation method thereof | |
| Eastoe et al. | The effect of nisin on the growth of cells and spores of Clostridium welchii in gelatine | |
| CN114671946B (en) | Recombinant human III-type collagen and preparation method and application thereof | |
| CN113063942B (en) | Indirect ELISA (enzyme-linked immuno sorbent assay) detection kit for detecting Morganella morganii antibody and application thereof | |
| CN116874570A (en) | Method for removing nucleic acid pollution in novel coronavirus nucleocapsid protein system |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230328 Address after: 361026 No. 188, Pingcheng South Road, Haicang street, Haicang District, Xiamen City, Fujian Province (third floor) Patentee after: Xiamen Baotai Biotechnology Co.,Ltd. Address before: 361000 No.188, Pingcheng South Road, Haicang street, Haicang District, Xiamen City, Fujian Province (3rd floor) Patentee before: Xiamen Baotai Biotechnology Co.,Ltd. Patentee before: Shenzhen wangrui Investment Co.,Ltd. |