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Chaperone/usher fusion proteins

A molecular chaperone, protein family technology, applied in the field of protein polymers, which can solve problems such as characteristics affecting nutrient diffusion, unsatisfactory three-dimensional framework, difficulty in generating specific cell lines or tissue type specificity, etc.

Inactive Publication Date: 2015-06-10
泰恩河上的纽卡斯尔大学
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current 3D skeleton is not ideal(7)
For example, it is difficult to generate scaffolds specific for a particular cell line or tissue type; there is also high batch-to-batch variability between these scaffolds; and changing a single property of these scaffolds without affecting their other properties is complicated
For example, if one wishes to alter the viscosity of a particular scaffold, one may end up altering both the density of the adhesion ligand and the presentation or porosity of the scaffold, which may affect the properties of nutrient diffusion through the scaffold (11)

Method used

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  • Chaperone/usher fusion proteins
  • Chaperone/usher fusion proteins
  • Chaperone/usher fusion proteins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0337] 1. Experiment overview

[0338] 1.1 Subcloning, expression and purification of Caf1

[0339]Using oligonucleotide primer pairs F1 forward (5′-ATA AAT CGG TTC AGT GGC CTC AAC GCT GTG-′3) and F1 reverse (5′-GGT TAG GCT CAA AGT AGG ATA ATT C-3′), Plasmid pAH34L(9) ((GenBank, accession number AY450847) encoding the caf operon as a template and KOD HOT START DNA polymerase (Novagen, UK) that produces blunt-ended fragments were used to pass PCR (30 amplification cycles; 95°C for 20 seconds, 55° C. for 10 seconds, 70° C. for 5 minutes) (PCR Express, Hybaid, UK) to amplify the caf operon (about 5.2 Kb in size). The obtained PCR product is loaded on a 0.7% agarose gel, and the bromine Ethidium (0.5 μg / Ml) was stained. DNA was visualized under a transilluminator (UV light at a wavelength of 254 nm) (Gel-Doc Bio-RAD, UK). Corresponding sections were excised from the agarose gel using sterile scalpel blades. The PCR product of the oligonucleotide of caf operon.Use QIA fast gel ex...

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Abstract

The invention provides achaperone / usher family polymer comprising at least one chaperone / usher family polypeptide monomer, wherein said at least one chaperone / usher family polypeptide monomer comprises an exogenous bioactive sequence.

Description

technical field [0001] The present invention relates to protein polymers comprising recombinant protein monomers with exogenous biologically active sequences. More particularly, the present invention relates to a chaperone / usher family polymer comprising at least one chaperone / usher family polypeptide monomer, wherein the at least one chaperone / usher family polypeptide monomer Contains exogenous biologically active sequences. Background technique [0002] In vivo proteins and other extracellular matrix (ECM) components form the interconnected mesh in which cells integrate and interact. One way to mimic this natural architecture is to create three-dimensional cell culture systems by cross-linking artificial polymers. [0003] The fields of cell culture and tissue engineering are well advanced and a variety of ECM equivalents have been developed. The materials used for the backbone differ among these equivalents, and thus the cell types that are able to propagate in them di...

Claims

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Application Information

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IPC IPC(8): A61K47/48C07K14/24C07K19/00
CPCA61L15/32A61L15/60A61L27/227A61L27/52A61L2300/424C07K14/24C07K2319/00C12N15/70
Inventor 杰里米·莱基
Owner 泰恩河上的纽卡斯尔大学
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