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Method of expressing thymys peptide alpha by yeast and use thereof

A technology of yeast expression and thymosin, applied in the fields of biology and genetic engineering, can solve the problems of high cost and high price, and achieve the effect of reducing production cost, avoiding pollution and saving foreign exchange.

Inactive Publication Date: 2004-02-04
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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  • Method of expressing thymys peptide alpha by yeast and use thereof
  • Method of expressing thymys peptide alpha by yeast and use thereof
  • Method of expressing thymys peptide alpha by yeast and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Method for expressing thymosin α1 in yeast:

[0023] 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 A 3', upstream primer 5' GGA AGC TTA CCA TGGGCT CTG ATG CTG CTG TTG 3', downstream primer 5' TAT CTA GAT TAT GGA TCCTAG TTC TCA 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 CatR6041 and BamH I endonuclease Cat R6021 were purchased from Pr...

Embodiment 2

[0031] Example 2 Study on Cellular Immunity and Humoral Immunity of Balb / c Mice Using Yeast Expressing Thymosin α1

[0032]Yeast-expressed thymosin α1 was used to gavage Balb / c mice to study the changes of their cellular and humoral immunity. It was confirmed that yeast-expressed thymosin α1 could increase the expression of IL-2, IFN-γ and their receptors in Balb / c mice. Expression level, has a powerful immune mediation effect.

Embodiment 3

[0033] Example 3 Application of Yeast Expressing Thymosin α1

[0034] The yeast-expressed thymosin α1 protein obtained by applying the method of the present invention is used for the treatment of duck hepatitis B animal models, and can significantly reduce the titer of hepatitis B virus; used for the treatment of rat liver tumors, can significantly reduce the size of tumors, and improve the efficiency of rat liver tumors. 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.

[0035] The thymosin α1 protein obtained by using the patented technology of this invention can be made into various preparations, such as beverages, syrups, etc., for the treatment of influenza, chronic hepatitis, low immune function, malignant tumors and other diseases, which has great social benefits and economic benefits.

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Abstract

A method for expressing the thymopeptide alpha 1 by yeast includes such steps as using PCR technique to amplify the whole-length gene of thymopeptide alpha 1, enzyme severing at both ends, and cloning it to yeast while stably expressing it in yeast. The expressed thymopeptide alpha 1 with very high bioactivity can be used to treat tumor, viral and immunodeficiency diseases, and hypothymism, or used as vaccine intensifier.

Description

technical field [0001] The invention belongs to biological technology, relates to genetic engineering, especially relates to a method for expressing thymosin α1 by yeast, and is suitable for adjuvant treatment of diseases caused by hypofunction of thymus. 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 c...

Claims

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Application Information

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IPC IPC(8): A61K38/32A61P31/12A61P35/00A61P37/04C12N15/16
Inventor 陈智陈峰
Owner ZHEJIANG UNIV PHARM CO LTD
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