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Temperature-sensitive polypeptide and coding gene thereof and preparation method and application thereof

An encoding gene and temperature-sensitive technology, applied in the field of temperature-sensitive polypeptides and their encoded genes, can solve problems such as poor synthesis and difficulty in obtaining temperature-sensitive polypeptides, and achieve the effects of high recovery efficiency, simple and fast development, and wide application range.

Active Publication Date: 2019-05-31
SHAANXI SCI TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First, it is difficult to obtain a temperature-sensitive polypeptide with a phase transition critical temperature suitable for protein separation and purification labels; second, a temperature-sensitive polypeptide is generally composed of multiple repeating units, which is not easy to synthesize

Method used

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  • Temperature-sensitive polypeptide and coding gene thereof and preparation method and application thereof
  • Temperature-sensitive polypeptide and coding gene thereof and preparation method and application thereof
  • Temperature-sensitive polypeptide and coding gene thereof and preparation method and application thereof

Examples

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Embodiment Construction

[0038] The principles and features of the present invention are described below in conjunction with the accompanying drawings, and the examples given are only used to explain the present invention, and are not intended to limit the scope of the present invention.

[0039] 1. Materials and Reagents

[0040] Restriction endonuclease, T4 polynucleotide kinase, calf intestinal alkaline phosphatase, and T4 DNA ligase were all purchased from NEB Company. DNA Marker was purchased from Nearshore Protein Technology Co., Ltd., and pre-stained protein Marker was purchased from Thermo Scientific. The plasmid extraction kit was purchased from OMEGA BIO-TEK, and the gel recovery kit was purchased from Beijing Broadtech Bio-Gene Technology Co., Ltd. The expression plasmid pET-28a, strain E.coli Top10 and BL21(DE3) are preserved by our laboratory.

[0041] 2. Design the monomeric gene of thermosensitive polypeptide

[0042] In the course of our research, we found that the polypeptide compo...

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Abstract

The invention relates to a temperature-sensitive polypeptide which comprises a plurality of short peptide repeating units. The sequence of each short peptide repeating unit is shown as SEQ ID NO.1. The invention also relates to a coding gene of the temperature-sensitive polypeptide. The invention also relates to a protein expression vector with the temperature-sensitive polypeptide coding gene asa purification label. The invention also relates to a preparation method of the protein expression vector. The invention also relates to a protein purification method. The synthesized temperature-sensitive polypeptide has the characteristic of reversible phase change, and the separation and purification of polypeptide molecules can be achieved through the reversible phase change circulation process. The temperature-sensitive polypeptide is used as a label to endow the fusion protein with reversible phase change property, and the protein separation and purification method is simple and fast todevelop, low in purification cost, high in recovery efficiency and wide in application range.

Description

technical field [0001] The invention relates to the field of protein separation and purification, more particularly, to a temperature-sensitive polypeptide and its coding gene, its preparation method and application. Background technique [0002] In the process of protein separation and purification, although chromatographic column chromatography is the most commonly used separation technique, this technique is time-consuming and labor-intensive, requires special separation equipment and reagents, and has high production costs. [0003] In order to simplify the separation operation steps, people began to explore different separation methods. The researchers found that some short peptides are temperature sensitive and undergo a reversible phase transition process as the ambient temperature changes. At a certain concentration, it is in a soluble state at low temperature; when the ambient temperature rises, the random coiled and stretched structure gradually forms a β-helix, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/00C12N15/11C12N15/70C12N15/66C12N15/62
Inventor 王珊珊
Owner SHAANXI SCI TECH UNIV
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