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Keratin BD-3, preparation method, pharmaceutical composition and application thereof

A technology of BD-3 and keratin, which is applied in the field of keratin BD-3, its preparation method and its pharmaceutical composition and application, to achieve the effects of high sample purity, prolonging the onset latency period, and strong effect

Active Publication Date: 2021-04-30
INST OF MATERIA MEDICA CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Using the protein expression system to prepare the target keratin, and then study its structure and function, no other literature reports, novel and creative

Method used

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  • Keratin BD-3, preparation method, pharmaceutical composition and application thereof
  • Keratin BD-3, preparation method, pharmaceutical composition and application thereof
  • Keratin BD-3, preparation method, pharmaceutical composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0114]Example 1 Shake flask fermentation Preparation Protein BD-3 Crude Solution A (TB Medization)

[0115]The nucleotide sequence shown in SEQ ID No. 2 is synthesized into a PET-28A (+) vector; sequencing determines the expression vector containing the correct sequence; transfection of the expression vector into BL21 (DE3) cells, The expression sensation-sensitive host cell containing the target nucleotide sequence was obtained. In the lB medium, in the shaker, it was cultured at 37 ° C, 220 rpm, and the recombinant strain was obtained.

[0116]The recombinant strain is pulled out in LBA flat line containing Kanamamycin, and the plate is inverted at a constant temperature incubator overnight for 16 hours overnight.

[0117]Configure 400 ml TB medium, 2 bottles, 200ml per bottle. Kanamamin (final concentration 50 μg / ml) was added to each bottle (200 mL), and a single column on the plate was added to Tb medium, in the shaker, overnight amplification of 37 ° C, 220 rpm. Got seed fluid.

[0118]...

Embodiment 2

[0123]Example 2 Shake flask fermentation Preparation Protein BD-3 Crude Solution B (Other Media)

[0124]The synthesis and sequencing determination of the expression vector containing the sequence shown in SEQ ID No. 2 is determined; the expression vector is transfected into BL21 (DE3) cells to obtain an expression sensation-sensitive host cell containing the target nucleotide sequence.

[0125]20 ml of LB medium was prepared, 800 μL was added to 50 μl of host cells containing the target coding sequence, in the shaker, under 37 ° C, 220 rpm, and cultured for 1 hour.

[0126]The above bacteria was taken in LBA plate contained in Kanamamycin, and the plate was inverted at a constant temperature incubator overnight for 16 hours in the 37 ° C.

[0127]10 ml of Lb medium was taken, and Kanamamicin (final concentration 50 μg / ml) was added, and a single bacterium on the plate was added to the LB medium, in the shaker, overnight amplification under 37 ° C, 220 rpm, and cultured for 15 hours, resultin...

Embodiment 3

[0133]Example 3 Fermentor Preparation Protein BD-3 Crude Solution C

[0134]The synthesis and sequencing determination of the expression vector containing the sequence shown in SEQ ID No. 2 is determined; the expression vector is transfected into BL21 (DE3) cells to obtain an expression sensation-sensitive host cell containing the target nucleotide sequence. In the lB medium, in the shaker, it was cultured at 37 ° C, 220 rpm, and the recombinant strain was obtained.

[0135]In LBA plates containing Kanamamicin, 100 μl of recombinant strains was added, and the coater was applied to uniform, and the plate was inverted at a constant temperature incubator overnight culture in a 37 ° C. Take three single collapses in a flat-panel in Kanamamycin, the plate overnight culture, and after three batches of shake flask fermentation expression verify that the strain was stored in 15% glycerol, it was separated into each 0.8ml, which was obtained. Working cell library, freezed to -80 ° C refrigerator s...

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Abstract

The invention relates to a keratin BD-3, a nucleic acid molecule encoding the keratin BD-3, an expression vector containing the nucleic acid molecule, a host cell containing the expression vector or a genome integrating the nucleic acid molecule, a preparation method of the keratin BD-3, and a pharmaceutical composition containing the keratin BD-3. The invention also discloses an application of the keratin BD-3, the nucleic acid molecule, the expression vector, the host cell or the pharmaceutical composition in preparation of antipyretic, analgesic, antitussive, expectorant, anticonvulsant, antiepileptic, antihypertensive, anti-inflammatory and antiviral drugs.

Description

Technical field[0001]The present invention relates to a keratin BD-3, a nucleic acid molecule encoding a cormope BD-3, an expression vector containing the nucleic acid molecule, and a host cell containing the expression vector or genome integrated the nucleic acid molecule, and keratin BD-3 The preparation method, a pharmaceutical composition containing such a keratin, and such a keratin and the pharmaceutical composition in preparing antipyretic analgesia, an antiplascent, anti-frial, anti-epileptic, lowering blood pressure, anti-inflammatory, anti-inflammatory Application in viral drugs.Background technique[0002]Keratin is a protein, widely existing in the epidermis of human and animals, and is the main component of hair, feather, hoof, shell, claw, corner, is an extremely important structural protein of connective tissue and plays a protective body. .[0003]Corrin is widely existed in the organism, is a renewable resource, with great utilization value, but has not been widely effi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12N15/12C12N15/70C12N1/21A61K38/17A61P29/00A61P11/14A61P11/10A61P25/08A61P9/12A61P31/12C12R1/19
CPCC07K14/4741C12N15/70A61P29/00A61P11/14A61P11/10A61P25/08A61P9/12A61P31/12A61K38/00A61P11/00C07K1/34A61K38/1748C07K14/78
Inventor 庾石山王晓良屈晶符江王玲蔡杰李密冯楠史国茹
Owner INST OF MATERIA MEDICA CHINESE ACAD OF MEDICAL SCI
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