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Mass production of high activity thymosin al by genetic engineering method

A technology of genetic engineering and thymosin, which is applied in the field of recombinant protein drug production, can solve problems such as difficult detection, low product activity, and high cost, and achieve the effects of reducing pollution, reducing production costs, and increasing production volume

Inactive Publication Date: 2006-10-04
BEIJING NORTHLAND BIOTECH
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  • Application Information

AI Technical Summary

Problems solved by technology

This preparation method has high requirements on raw material purity, large investment in synthetic instruments and equipment, complex production process (synthesis and chemical modification, etc.), low product activity, high cost and other deficiencies; genetic engineering methods use bioengineering technology to construct thymosin α1 Gene expression The vector is expressed in Escherichia coli and produced by separation and purification. The product obtained has high purity and activity. However, since the small peptide molecule is easily degraded and difficult to be detected after expression, its yield and yield cannot reach industrialization. Require

Method used

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  • Mass production of high activity thymosin al by genetic engineering method
  • Mass production of high activity thymosin al by genetic engineering method
  • Mass production of high activity thymosin al by genetic engineering method

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Experimental program
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Effect test

Embodiment Construction

[0010] 1. Construction of thymosin a1 precursor protein expression plasmid and acquisition of engineering bacteria

[0011] Firstly, the whole gene sequence of the thymosin a1 precursor protein was amplified through the whole gene synthesis, and the design was as follows:

[0012] GGA TCC CCG CGT GCG TCG GAT GCG GCC GTG GAT

[0013] ACC TCG TCG GAA ATT ACC ACG AAA GAT CTG AAA

[0014] GAA AAA AAG GAA GTG GTT GAA GAG GCG GAA AAT

[0015] TAA TGA CTC GAG

[0016] Using the GST gene as a template, a product of about 500 bp was amplified by PCR. After sequencing plasmid cloning and gene sequencing, the designed sequence was obtained for future use.

[0017] The target fragments were digested by BamH I, EcoR I, and XhoI to recover small fragments respectively, and the plasmid PGM was digested by XhoI and EcoRI to recover large fragments, and the three target gene fragments were ligated in the T4 ligase system at 18°C. The recombinant plasmid obtained by screening was named PGMT...

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Abstract

The related fused protein prepared by both of one protein stable expressed in bacterium and the natural thymosin a1 can take large-production soluble expression in colibacillus. It also relates a1 derivant with high biological activity. This invention can be used on assisted cancer cure.

Description

1. Technical field [0001] The invention belongs to the field of recombinant protein drug production. Through the genetic engineering construction of the precursor fusion protein, the high-efficiency expression of the target protein is realized. This method has the characteristics of high expression, easy operation, and high activity of the mature protein. 2. Background technology [0002] The immune system is the body's defense system. The main cells involved in the immune response are T lymphocytes, B lymphocytes and macrophages. The thymus is the central immune organ for the development and differentiation of T lymphocytes. Thymus factors (or hormones) secreted by the thymus are a series of essential substances required for the development and differentiation of T lymphocytes, among which thymosin α1 (Tα1) is a major thymus factor whose structure and function have been clarified. [0003] Tα1 is a polypeptide isolated from thymosin fraction 5 (TF5) first extracted from ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/02C12N15/09C12N15/63C12N15/70C12N15/81
Inventor 许松山宣锺健马素永聂李亚
Owner BEIJING NORTHLAND BIOTECH
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