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Recombination human ciliary neurotrophy factor, mutant and application thereof

A ciliary neurotrophic and mutant technology, which is applied in the fields of application, drug combination, and genetic engineering, can solve the problems of poor stability and low expression efficiency of expressed proteins, and achieve enhanced biological activity, increased stability, and easy purification and complexation. sexual effect

Active Publication Date: 2008-03-19
LANZHOU INST OF BIOLOGICAL PROD
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Problems solved by technology

However, the efficiency of directly expressing the natural human ciliary neurotrophic factor gene in Escherichia coli is low, and the stability of the expressed protein is relatively poor

Method used

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  • Recombination human ciliary neurotrophy factor, mutant and application thereof
  • Recombination human ciliary neurotrophy factor, mutant and application thereof
  • Recombination human ciliary neurotrophy factor, mutant and application thereof

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specific Embodiment approach

[0018] The whole gene synthesis of the recombinant CNTF of the present invention is completed by Shanghai Sangon Bioengineering Technology Service Co., Ltd. When performing gene synthesis, it is required to design two enzyme cutting sites, NdeI and AvrII, respectively, at the upper and lower ends of the target gene. The synthesized target gene is 603bp in full length, loaded into the cloning vector PUC18 plasmid, the sequenced correct target gene is cut out, inserted into the expression vector pETr, and the protein is expressed and sequenced.

[0019] Compared with the natural human CNTF gene, the cDNA synthesized by the present invention replaces the cysteine ​​at the 17th position of the human ciliary neurotrophic factor gene with alanine, and the glutamic acid at the 63rd position with arginine At the same time, part of the codons in the original CNTF gene were changed to codons preferred by prokaryotes, but no amino acid changes were involved, which was a synonymous mutatio...

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Abstract

The present invention relates to a recombinant human ciliary nerve nutrition factor, a mutant of the nutrition factor, and the preparation and the application thereof. The present invention provides the recombinant human ciliary nerve nutrition factor that the 17th thioserine of a natural CNTF gene is changed into alanine, the 63rd glutamic acid of the protogene is changed into arginine, at the same time, the partial codons in the CNTF protogene are changed into the codons which is favored by prokaryotes, but the code of the gene is not changed. On the basis, the recombinant CNTF is modified, to form different mutants. The mutants of the present invention has better effect on the aspect of curing the ordinary obesity or diabetes type obesity.

Description

technical field [0001] The present invention relates to recombinant human ciliary neurotrophic factor, mutants of the trophic factor, and their preparation and application. Background technique [0002] Ciliary neurotrophic factor (CNTF) is a neurocytokine, expressed in the peripheral and central nervous system since the late embryonic period, its molecular weight is 22.75KDa, including 200 amino acids. It has been disclosed in the literature that recombinant human ciliary neurotrophic factor (CNTF) is used to treat common obesity and diabetic obesity. Such as: Chinese invention patent 200410030756 discloses the treatment of obesity with CNTF and its derivatives; Chinese invention patent 97180757 discloses that hCNTF and its mutants or other molecules that activate CNTF receptors are used for the treatment of obesity and related diseases (such as hyperglycemia) medicinal use. However, the efficiency of directly expressing the natural human ciliary neurotrophic factor gene ...

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Application Information

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IPC IPC(8): C12N15/12C07K14/435A61K38/17A61P3/04A61P3/10
Inventor 蒋琳应莲芳雷清谢溱梁凌宇马亚茹辛芳杨军
Owner LANZHOU INST OF BIOLOGICAL PROD
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