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Preparation method of asymmetric virus nanoparticles

A nanoparticle and inorganic nanoparticle technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, chemical instruments and methods, etc., to achieve the effect of convenient mass acquisition, easy control of reaction conditions, and large-scale production.

Inactive Publication Date: 2012-01-18
SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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  • Claims
  • Application Information

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Problems solved by technology

However, manipulating their self-assembly in a controllable way at the molecular level and accurately analyzing their assembly products remain challenging tasks

Method used

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  • Preparation method of asymmetric virus nanoparticles
  • Preparation method of asymmetric virus nanoparticles
  • Preparation method of asymmetric virus nanoparticles

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Embodiment 1

[0042] Introduce Cys into the loop structure of the main capsid protein vp1 of SV40 (that is, the 74th amino acid is mutated to Cys), insert the His-tag at the 139th position, and construct the vector PET32a-His-mvp1 (His-mvp1); confirmed by sequencing The correctness of the target gene sequence. Transform PET32a-His-mvp1 into E. coli For Rosetta (DE3) competent cells, single clones were picked from the plate 12 hours after plating and transferred to 5 mL LB test tube culture medium, added ampicillin and chloramphenicol, and incubated overnight at 37°C and 180 r / min. According to the 1% inoculum amount, transfer to 5 ml LB test tube culture medium (3 parallel tubes), add corresponding antibiotics, and culture at a constant temperature of 37°C and 180 r / min for about 2.5 h (OD600 between 0.4 and 0.6). One of the tubes was used as a blank control (without IPTG), and the other two tubes were added with IPTG at a final concentration of 1 mM inducer. After induction and culture a...

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Abstract

The invention discloses a preparation method of asymmetric virus nanoparticles. The method comprises the following steps: carrying out gene modification on the virus capsid protein surface, so that the virus capsid protein simultaneously has coupled functional group and separate group; thoroughly mixing the modified virus capsid protein and wild type virus capsid protein while controlling the proportion of the two virus capsid proteins; and meanwhile, adding corresponding inorganic nanoparticles according to the total amount of the virus capsid protein to implement controllable assembly of the virus nanoparticles, thereby obtaining the asymmetric functionalized nanoparticles which are the goal product. The invention adopts biomacromolecule-protein as the nano material, and the biomacromolecule-protein can be easily modified and manually operated, and can be conveniently obtained massively. On the basis of the structural symmetry of the self-assemblable virus capsid protein, the two different protein molecules can be assembled in an oriented mode according to the previous design, and therefore, the assembly body has diversity and controllability; and the invention has the advantage of manageable reaction conditions, and can implement large-scale production.

Description

technical field [0001] The invention relates to a controllable method at the molecular level to manipulate the self-assembly of biomacromolecules, in particular to a preparation method of asymmetric virus nanoparticles (asymmetric VNPs), which belongs to the field of nano-biotechnology. Background technique [0002] Nanotechnology refers to the technology of studying the laws and characteristics of electrons, atoms and molecules, and processing substances and materials on a scale of 0.1 to 100 nanometers. In 1981, scientists invented the scanning tunneling microscope, an important tool for nanometer research, and the world of atoms and molecules can be seen from then on. In 1990, the first international nanotechnology conference was held in Baltimore, USA, and the form of nanotechnology was born. [0003] Due to their unique optical, electrical, magnetic and mechanical properties, and because of their small size, large specific surface area, high surface energy, and large s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/04C12N7/02C12N15/37C12N15/70C07K14/025
Inventor 王强斌李峰陈艳华
Owner SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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