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Method for expressing and purifying protein by using calsequestrin-tag

A protein and labeling technology, which is applied in the field of protein expression and purification using troponin labeling, can solve problems such as difficulty in obtaining target proteins, achieve the effect of reducing protein material and drug costs, and easy separation and purification

Pending Publication Date: 2022-02-11
KOREA INST OF CERAMIC ENG & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This task of obtaining target proteins from transformed cells is not easy

Method used

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  • Method for expressing and purifying protein by using calsequestrin-tag
  • Method for expressing and purifying protein by using calsequestrin-tag
  • Method for expressing and purifying protein by using calsequestrin-tag

Examples

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Effect test

no. 1 example

[0072] According to a first example, the present invention provides a fusion protein comprising a troponin tag and a target protein.

[0073] The present invention is characterized in that, in the fusion protein of the present invention, the troponin tag includes troponin 1 (CSQ1) or troponin 2 (CSQ2).

[0074] The present invention is characterized in that, in the fusion protein of the present invention, the above-mentioned troponin 1 and troponin 2 are respectively encoded by amino acids consisting of SEQ ID NO:1 and SEQ ID NO:2. The above-mentioned amino acid of SEQ ID NO:1 can be encoded by the nucleotide sequence of SEQ ID NO:3, and the above-mentioned amino acid of SEQ ID NO:2 can be encoded by the nucleotide sequence of SEQ ID NO:4.

[0075] The present invention is characterized in that, in the fusion protein of the present invention, the above-mentioned troponin tag and the target protein are fused by a hydrolase decomposing peptide composed of amino acids, and the ab...

no. 3 example

[0083] According to a third embodiment, the present invention provides cells transformed with an expression vector comprising a fusion protein comprising a troponin tag and a target protein.

[0084] The present invention is characterized in that, in the transformed cell of the present invention, the above-mentioned troponin tag is encoded by an amino acid consisting of SEQ ID NO:1 or SEQ ID NO:2. The above-mentioned amino acid of SEQ ID NO:1 can be encoded by the nucleotide sequence of SEQ ID NO:3, and the above-mentioned amino acid of SEQ ID NO:2 can be encoded by the nucleotide sequence of SEQ ID NO:4.

[0085] The present invention is characterized in that, in the transformed cells of the present invention, the above-mentioned troponin tag and target protein can be fused by a hydrolase composed of amino acids selected from the group consisting of SEQ ID NO:5 to SEQ ID NO:8 composed of groups.

[0086]The present invention is characterized in that, in the transformed cells...

Embodiment approach

[0095] Hereinafter, in order to facilitate the understanding of the invention, various examples are presented. The following examples are only provided for easier understanding of the invention, and the protection scope of the invention is not limited to the following examples.

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Abstract

The present invention relates to a method for expressing, water-solubilizing, and purifying protein by using a calsequestrin-tag (CSQ-tag). Provided are: a recombinant expression vector containing a CSQ-tag and a target protein; a host cell transformed using the recombinant expression vector; and a method for expressing and purifying a target protein by using the CSQ tag. The present invention uses CSQ-tags to increase the expression of proteins that are widely used in pharmaceuticals and cosmetics, and allows the proteins to be easily separated by calcium, and thus is expected to be able to lower the cost of protein materials and protein pharmaceuticals.

Description

technical field [0001] The present invention relates to methods for expressing, dissolving and purifying a protein using a calsequestrin-tag (calsequestrin-tag), providing a fusion protein comprising a calsequestrin-tag and a target protein, including nucleosides encoding the above-mentioned fusion protein The nucleic acid of the acid sequence, the expression vector comprising the above nucleic acid, the cell transformed by the above expression vector and the expression, water solubilization and purification method of the target protein using the troponin tag. The present invention uses the calsequestrin label to increase the expression and water solubility of proteins widely used in medicines and cosmetics, and makes the proteins easily separated by calcium, thereby reducing the cost of protein materials and medicines. Background technique [0002] Recently, with the development of genetic engineering and biology, many attempts have been made to mass-produce or obtain speci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435
CPCC07K14/485C07K2319/20C07K14/4728C07K14/49C07K14/495C07K14/50C07K14/51C07K14/605C07K14/78C07K2319/50C12N9/0065C12N9/6429C12N9/6472C12Y111/01007C12Y304/21005C12Y304/22044
Inventor 金成炫方珍浩崔美贤
Owner KOREA INST OF CERAMIC ENG & TECH
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