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Production of high-activity extrasin beta 4 derivative by gene engineering

A technology of derivatives and thymosin, which is applied in the production of recombinant protein drugs and derivatives, can solve the problems of production rate and yield not meeting industrialization requirements, high raw material purity requirements, and low product activity.

Inactive Publication Date: 2007-01-17
BEIJING NORTHLAND BIOTECH
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AI Technical Summary

Problems solved by technology

This preparation method requires high purity of raw materials, large investment in synthetic instruments and equipment, complicated production process (synthesis and chemical modification, etc.), low product activity, and high production cost.
The conventional genetic engineering method constructs thymosin β4 gene expression vector, expresses it in Escherichia coli, and makes it through separation and purification. The obtained product has high purity and activity. Its yield and yield all can't reach industrialization requirement

Method used

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  • Production of high-activity extrasin beta 4 derivative by gene engineering
  • Production of high-activity extrasin beta 4 derivative by gene engineering
  • Production of high-activity extrasin beta 4 derivative by gene engineering

Examples

Experimental program
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Effect test

Embodiment Construction

[0014] 1. Construction of thymosin β4 precursor protein expression plasmid and acquisition of engineering bacteria:

[0015] First, the whole gene sequence of thymosin β4 precursor protein was amplified by whole gene synthesis:

[0016] GGA TCC CCT CGA GCT TCT GAC AAA CCC GAT ATG GCT GAG ATC GAG AAA TTC

[0017] GAT AAG TCG AAA CTG AAG AAG ACA GAG ACG CAA GAG AAA AAT CCA CTG CCT

[0018] TCC AAA GAA ACG ATT GAA CAG GAG AAG CAA GCA GGC GAA TCG TAA GTC GAC

[0019] 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.

[0020] 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 T4 ligase system at 18°C. The recombinant plasmid obtained by screening was named PGMTβ4....

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Abstract

A high-activity extrasin beta-4 derivative by gene engineering method is prepared by expressing in bacterium, combing protein with natural extrasin beta-4, dissolved expressing in colibacillus, cutting and processing by fusion protease. It has stable and efficient expression, higher purity and can be used to repair skin and heart damage.

Description

1. Technical field [0001] The invention belongs to the field of recombinant protein drug production and derivative 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 purity of the final product Ala-Tβ4 (greater than 98%). 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 β4 (Tβ4) is a major thymus factor whose structure and function are relatively clear. As shown in a study using the activity o...

Claims

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Application Information

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IPC IPC(8): C07K14/435C07K14/47C12N15/62A61K38/17A61P17/00A61P9/10A61P27/02
Inventor 聂李亚马素永许松山文美玉
Owner BEIJING NORTHLAND BIOTECH
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