Ferritin fusion protein with galactose binding lectin EW29 tag, protein cage nanoparticle and preparation method of protein cage nanoparticle

Abstract

The invention discloses ferritin fusion protein with a galactose binding lectin EW29 tag, a protein cage nanoparticle and a preparation method of the protein cage nanoparticle. Ferritin is linked withthe EW29 tag and a TEV protease cutting site to obtain a EW29 tag / TEV protease recognition site / ferritin recombinant sequence, and the fusion protein is obtained after induction and affinity purification. Electronic speculum results show that the fusion protein forms the protein cage nanoparticle of 20-25 nm. The constructed EW29 / ferritin vector can be used as a novel dissolution, purification and encapsulation platform and can serially link different target proteins in the ferritin so as to improve solubility and sensitivity of the target proteins.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a ferritin fusion protein with a galactose-binding lectin EW29 label, protein cage nanoparticles and a preparation method thereof. Background technique [0002] Through DNA recombination technology, fusion protein has been easily expressed in prokaryotic expression system (Escherichia coli) and eukaryotic expression system (yeast and mammalian cells), and its products have been widely used in biology and medicine. Rapid development. Compared with eukaryotic expression systems, Escherichia coli is still the main host for recombinant protein production due to its advantages of easy operation, low cost and high yield. However, when expressing recombinant proteins in the E. coli bacterial system, there are many shortcomings that are difficult to overcome: 1. Recombinant proteins often appear in the form of inactive inclusion bodies; 2. Lack of eukaryotic post-translational m...

AI Technical Summary

Problems solved by technology

However, when expressing recombinant proteins in the E. coli bacterial system, there are many shortcomings that are difficult to overcome: 1. The recombinant proteins often appear in the form of inactive inclusion bodies; 2. Lack of eukaryotic post-translational modifications (glycosylation, phosphorylation) and acetylation, etc.) mechanism, although the obtained recombinant protein is correct in the primary amino acid sequence, it is quite different from the natural protein in the advanced structure and conformation, and has no activity or very poor activity; 3. Host cell (Escherichia coli) own protein Becoming a pyrogen, difficult to remove, and safety, these problems limit the further application of prokaryotic expressed recombinant proteins in practice

For affinity tags, 6×His is often chosen, but imidazole molecules are introduced during the purification process of conventional His affinity tags, which has toxic effects, and imidazole residues may still affect the protein after desalting, causing damage to the animal body; usually in order to increase the concentration of the target protein Soluble, also requires an additional tandem solubilizing tag

Images