Metal affinity fusion protein tag and its application

Abstract

The invention discloses a novel metal affinity fusion protein tag, which is derived from an extracellular amino terminal fragment of a natural human copper ion transporter, which can be used as the metal affinity tag instead of His-tag, and can be used for purifying a recombinant fusion protein; the invention also discloses a novel fusion protein fused to the tag, and the novel fusion protein hasparticular physiological and pharmacological activities.

Description

technical field [0001] The invention belongs to the field of bioengineering, and specifically relates to a novel metal-affinity fusion protein tag having affinity for metal ions, and a fusion protein thereof. Background technique [0002] Metal chelate affinity chromatography, also known as immobilized metal ion affinity chromatography. This method utilizes the metal ion binding properties of some amino acids on the protein surface, such as histidine, tryptophan and other residues, to separate and purify proteins through the adsorption of solid-phase substrates. Histidine tag (His-Tag), as a synthetic sequence, can be used for the fusion expression of foreign proteins to facilitate the purification of recombinant proteins. A peptide composed of continuous histidine residues (usually 6-10 histidines) is fused to the amino-terminal, carboxy-terminal or middle of the target protein, which can be expressed in various expression systems (bacteria, yeast and mammalian cells) suc...

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Problems solved by technology

[0003] It has been found that the fusion of His-tag at the amino or carboxyl terminal of the protein will affect the renaturation[1], stability[2] or solubility[3] of the protein; His-tag may also affect the activity of the protein, such as His -tag is fused to the amino terminal of Chlorocatchol 1,2-Dioxygenase (CCD), although its structure is only slightly different from CCD, its activity is reduced by 5 times[4]

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