Method for expressing thymopolypeptides šŒ1 by use of escherichia coli and its application

A technology of Escherichia coli and thymosin, which is applied in the fields of biology and genetic engineering, can solve the problems of high price and high cost, and achieve the effects of low development cost, low production cost and good biological activity

Inactive Publication Date: 2003-07-23
ZHEJIANG UNIV PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to its high cost and high price, the content of each Ridaxian is 1.6mg, the price is more than 800 yuan, and a course of treatment is more than 50,000 yuan, which is unbearable for ordinary patients.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 The method for expressing thymosin α1 in Escherichia coli:

[0019] 1. Acquisition of full-length thymosin α1 gene: synthetic full-length thymosin α1 single-stranded DNA, sequence: 5'AAC GGA ATT CAT GTC TGA TGC TGC TGT TGA TAC TTC TTC TGA GATTAC TAC TAA AGA TCT TAA GGA GAA GAA GGA AGT TGT CGA AGA GGCTGA GAA CTA GGA TCC ATA A3', upstream primer 5' GGA AGC TTA CCA TGG GCTCTG ATG CTG CTG TTG 3', downstream primer 5' TAT CTA GAT TAT GGA TCC TAG TTCTCA GC 3'. PCR reaction amplification: 94°C for 30 seconds, 54°C for 30 seconds, 72°C for 30 seconds, a total of 30 cycles, to obtain thymosin α1 full-length double-stranded DNA, with HindIII restriction site upstream and BamH I restriction site downstream . (Thymosin α1 single-stranded DNA and upstream and downstream primers were synthesized by Shanghai Boster Company, high-fidelity Pwo DNA polymerase was purchased from Roche Company, Cat.1644947; HindIII endonuclease Cat R6041 and BamH I endonuclease Cat R6021 were pu...

Embodiment 2

[0026] Example 2 Study on Cellular Immunity and Humoral Immunity of Balb / c Mice Using Escherichia coli to Express Thymosin α1

[0027] Thymosin α1 expressed in Escherichia coli was injected intramuscularly into the thighs of Balb / c mice to study the changes in cellular and humoral immunity. The expression level of the receptor has a powerful immune mediating effect.

Embodiment 3

[0028] Example 3 Application of Escherichia coli to express thymosin α1

[0029] The thymosin α1 protein expressed by the Escherichia coli obtained by the method of the present invention is used for the treatment of duck hepatitis B animal model, and can significantly reduce the titer of duck hepatitis B virus; it is used for the treatment of rat liver tumors, can significantly reduce the tumor size, improve Rat cellular immunity level; for immunocompromised mice, such as mice treated with cyclophosphamide, thymosin α1 can significantly improve their immune function, serum levels of IL-2, IFN-γ and their receptors.

[0030] The thymosin α1 protein obtained by using the patented technology of the present invention can be made into injection preparations for the treatment of influenza, chronic hepatitis, low immune function, malignant tumors and other diseases, which has great social and economic benefits.

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PUM

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Abstract

A process for expressing thymopeptide alpha 1 by colibacillus includes PCR amplification to obtain the full-length gene of thymopeptide alpha 1, endonuclease severing at both ends, and cloning to colibacillus for stable expression. After purified, it can be used as injection to treat immunodeficiency diseases.

Description

technical field [0001] The invention belongs to biotechnology and relates to genetic engineering, especially a method for expressing thymosin α1 in Escherichia coli, which can be used for adjuvant treatment of diseases with low immune function. Background technique [0002] The thymus is an important immune organ in the body and plays an important role in the development of the lymphatic system and maintaining the normal function of the immune system. Especially in anti-infection, anti-tumor, transplantation and autoimmune diseases, it has special significance. As the body ages, the thymus gradually shrinks and degenerates, and the incidence of tumors, infections, and immune diseases gradually increases. A large number of studies have shown that the calf thymus extract (containing thymosin) can be used as an adjuvant treatment for tumors, infections and other diseases caused by hypofunction of the thymus. However, since the thymus extract of animals is a mixed component, t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/16A61K39/39A61P31/12A61P35/00C12N15/70C12P19/34C12P21/02
Inventor 陈智陈峰
Owner ZHEJIANG UNIV PHARM CO LTD
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