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Small peptide gene for resisting rheumatoid arthritis and production method of small peptide gene

A production method and arthritic technology, applied in the field of anti-rheumatoid arthritis small peptide gene and its production, to achieve the effects of reducing pollution, facilitating industrialization and low cost

Inactive Publication Date: 2015-01-28
BEIJING SHENGKE DONGFANG TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The technical problem to be solved by the present invention is: the anti-rheumatoid arthritis small peptide dnaJP1 is a short peptide composed of several amino acids, and it is very difficult to express and isolate the short peptide directly in the host bacteria, and adopt fusion expression The method also has very low expression due to the use of larger fusion tags and lower copies of anti-inflammatory genes

Method used

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  • Small peptide gene for resisting rheumatoid arthritis and production method of small peptide gene
  • Small peptide gene for resisting rheumatoid arthritis and production method of small peptide gene
  • Small peptide gene for resisting rheumatoid arthritis and production method of small peptide gene

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Embodiment Construction

[0021] Construction of engineering bacteria:

[0022] The content includes designing the sequence according to the codon preference, synthesizing the whole gene sequence, digesting the plasmid with Alwn I, introducing the target sequence, and constructing the E. coli BL21(DE3) pLysS with pet-31(b+) as the expression vector and the host as Bacterial MDNAJP1 fusion expression system. Amino acid sequence of the six repeats of MDNAJP1, according to E. coli codon bias. The determined gene sequence is shown in SEQ ID NO:1.

[0023] Gene sequences were synthesized by our own laboratory. The plasmid was cut with AlwnI endonuclease, the target gene fragment was subcloned, ligated into the plasmid, and then transformed into Escherichia coli. Both the host bacteria and the expression vector were purchased from Novagen. After the construction of the engineering bacteria was completed, the expression plasmid was digested and identified, and the DNA sequencing of the target fragment was ...

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Abstract

The invention discloses a small peptide gene for resisting rheumatoid arthritis, and the nucleotide sequence is indicated as SEQIDNO.1. The derivative M-DNAJP1 of the DNAJP1 is produced by utilizing a gene recombination method, the gene of the dnaJP1 is artificially established, and M-dnajp1-M-dnajp1-M-dnajp1-M-dnajp1-M-dnajp1-M-dnajp1 is formed in a series connection manner by virtue of a method for connecting in an end-to-end manner and adding methionine, cloned into plasmid pet31b(+), then converted to enter colibacillus to be expressed to obtain the DNAJP1 modified polypeptide, and the repeated sequence of the M-DNAJP1 is indicated as SEQIDNO.2. The small peptide is prepared in a fermenting manner, the free polypeptide is obtained by utilizing a cyanogen bromide dissociation method, the dissociated cyanogen bromide is processed at a high temperature and high pressure to generate ammonia water and sodium formate, the environmental pollution is reduced, the cost is low in the entire process, the yield is high, simplicity in purification is realized, and the industrialization is facilitated.

Description

technical field [0001] The invention relates to the recombinant fermentation production and purification method of a small anti-rheumatoid arthritis peptide. Background technique [0002] Rheumatoid arthritis is a chronic and recurrent painful disease mainly caused by systemic joint inflammation. The English name Rheumatoid arthritis, abbreviated as RA, is a common and frequently-occurring disease. The onset time can last for several days, weeks or months, and have different degrees of activity. In severe cases, patients cannot take care of themselves, often involving their whole life, forming long-term illnesses, which greatly endanger human life and health. People expect It can develop and produce more efficient and high-quality drugs for clinical application. [0003] Currently applied RA treatment drugs include anti-inflammatory drugs, such as aspirin, corticosteroids, and drugs that reduce disease symptoms by suppressing or eliminating the body’s immune response. Altho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K14/47C12N15/70C12P21/02
Inventor 李相鲁张严冬孟建军梁庄严
Owner BEIJING SHENGKE DONGFANG TECH
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