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Multifunctional composite supermolecule nanofiber self-assembly system and application thereof

A nanofiber and supramolecular technology, applied in the field of protein self-assembly, can solve the problems of discount of practical value and neglect of functionality

Inactive Publication Date: 2017-12-08
SHANGHAI TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the nanofibrous structure prepared by the above methods can not take into account the unity of structure and function. It often realizes the diversification of structure but ignores its functionality as a material, so its practical value is great. discount

Method used

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  • Multifunctional composite supermolecule nanofiber self-assembly system and application thereof
  • Multifunctional composite supermolecule nanofiber self-assembly system and application thereof
  • Multifunctional composite supermolecule nanofiber self-assembly system and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] Example 1: Construction, expression and purification of functional FUS LC protein

[0098] The R-FUS LC expression plasmid was constructed, and the R-FUS LC expression plasmid was transformed into BL21(DE3) Escherichia coli to express the R-FUS LC fusion protein, and R is a functional protein.

[0099] The present invention utilizes a series of fusion plasmids expressing R-FUS LC, and combines peptide proteins with different functional groups. The plasmid map is as follows: figure 2 shown. Among them, pHis-FUS LC, pHis-mCherry-FUS LC, pHis-CFP-FUSLC and pHis-EGFP-FUS LC plasmids were donated by Professor McKnight's research group at Southwestern Medical College, USA. The amino acid sequence of FUS LC is shown in SEQ ID NO: 1 , the amino acid sequence of EGFP is shown in SEQ ID NO: 2, the amino acid sequence of mCherry is shown in SEQ ID NO: 3, the amino acid sequence of CFP is shown in SEQ ID NO: 4, the amino acid sequence of Histag is shown in SEQ ID NO: 10, and pHis...

Embodiment 2

[0124] Example 2: Preparation and characterization of single-component recombinant FUS LC nanofibers

[0125] A multifunctional composite supramolecular nanofiber self-assembly system, comprising pHis-mMaple3-FUS LC obtained in Example 1, pHis-PAtagRFP-FUS LC, pHis-EGFP-FUS LC-Spycatcher, pHis-EGFP-FUS LC- One of mefp5, pHis-FUS LC, pHis-mCherry-FUS LC, pHis-CFP-FUS LC and pHis-EGFP-FUSLC proteins.

[0126] The self-assembly system of multifunctional composite supramolecular nanofibers was used to prepare single-component recombinant FUS LC nanofibers:

[0127]The target protein obtained in Example 1 (pHis-mMaple3-FUS LC, pHis-PAtagRFP-FUS LC, pHis-EGFP-FUS LC-Spycatcher, pHis-EGFP-FUS LC-mefp5, pHis-FUS LC, pHis-mCherry -FUS LC, pHis-CFP-FUS LC, and pHis-EGFP-FUS LC) were concentrated to 40 μM, and left at 4°C (or room temperature) overnight to obtain a large number of supramolecular nanoparticles produced by the self-assembly of the target protein fiber, but there are stil...

Embodiment 3

[0139] Example 3: Preparation and Characterization of Bicomponent Random Copolymer Recombined FUS LC Nanofibers

[0140] A multifunctional composite supramolecular nanofiber self-assembly system, comprising the pHis-EGFP-FUS LC and pHis-mCherry-FUS LC proteins obtained in Example 1.

[0141] In view of the strong fiber growth and gel formation ability of the pHis-EGFP-FUS LC and pHis-mCherry-FUS LC proteins demonstrated in Example 2, the pHis-EGFP obtained in Example 1 with the same concentration of the substance (40 μM) -FUS LC and pHis-mCherry-FUS LC protein monomer solution were mixed together according to the ratio of 1:3, 1:1 and 3:1 to form the multifunctional composite supramolecular nanofiber self-assembly system and left at 4°C ( or dialyzed overnight) to self-assemble into recombinant FUSLC nanofibers ( Figure 9 a, 9b, 9c). Upright fluorescence microscope characterization as Figure 9 As shown in d, it can be proved that with the expansion of the component ratio ...

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Abstract

The invention provides a multifunctional composite supermolecule nanofiber self-assembly system and application thereof. The multifunctional composite supermolecule nanofiber self-assembly system is characterized by comprising at least one FUS LC combined with a functional protein R. The FUS LC can be combined with various functional proteins through gene engineering without influencing the capacity of assembling fiber, and the amyloid protein can realize structure controllability and functional diversification through slow fiber growth capacity and diversified functional proteins.

Description

technical field [0001] The invention relates to the fields of protein self-assembly, genetic engineering technology and multifunctional supramolecular materials, in particular to a method and application for making multifunctional composite materials with complex structures by using a self-assembled protein molecule. Background technique [0002] Supramolecules generally refer to a class of substances that use weak non-covalent interactions such as hydrogen bonds, hydrophobic forces, π-π conjugation, and van der Waals forces to gather a large number of monomers together and maintain a relatively stable structure. 1 . Existing widely in nature, functionalized one-dimensional nanofiber materials are formed by regulating the self-assembly of discrete monomers, such as microtubules and muscle fibers that maintain cell survival and assist cell movement in life, that is, through the weak inter-monomer Hydrogen bonding forces, self-assembled to form one-dimensional nanofibrous mat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70B82Y30/00B82Y40/00
CPCC12N15/70B82Y30/00B82Y40/00C07K14/47C07K2319/60C07K2319/735C07K2319/21
Inventor 钟超安柏霖崔孟奎
Owner SHANGHAI TECH UNIV
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