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Stable coronavirus recombinant protein dimer and its expression vector

A coronavirus and recombinant protein technology, applied in the field of recombinant protein, can solve the problems of poor protein stability and weak immunogenicity, achieve good immunogenicity, avoid degradation, and improve immunogenicity

Active Publication Date: 2021-03-26
SOUTH CHINA UNITED VACCINE INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when the S-RBD monomer is used as an antigen, the immunogenicity is weak and the protein stability is poor

Method used

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  • Stable coronavirus recombinant protein dimer and its expression vector
  • Stable coronavirus recombinant protein dimer and its expression vector
  • Stable coronavirus recombinant protein dimer and its expression vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1: A kind of vector expressing novel coronavirus SARS-CoV-2 recombinant protein

[0060] Schematic diagram of the recombinant protein structure of SARS-CoV-2 figure 1 shown.

[0061] The N-terminal signal peptide MFVFLVLLPLVSS of the new coronavirus SARS-CoV-2 S protein is connected to the N-terminal of SARS-CoV-2 S-RBD, and the CTD region (aa248-365) of the N protein is connected to the C-terminal of the S-RBD. Use the linker connection in the middle. placed in pFastBac TM P of the SCUVI plasmid polh under the promoter. Obtain the shuttle vector pFastBac TM SCUVI-SP-S-RBD-CTD is a vector expressing recombinant protein of novel coronavirus SARS-CoV-2.

Embodiment 2

[0063] The preparation method of the vector expressing novel coronavirus SARS-CoV-2 recombinant protein, comprises the following steps:

[0064] 1) Gene optimization synthesis

[0065] Link the N-terminal signal peptide SP, 306-575 region (S-RBD) of the new coronavirus SARS-CoV-2 S protein with the CTD region (aa248-365) of the N protein (S-RBD and CTD are connected with Linker GSGSG), According to the host cell Spodoptera frugiperda Codon source optimization for the preference of Spodoptera frugiperda (Sf9) cell line, PUC57 as the vector, and xho I , KpnI Synthesize SP-S-RBD-CTD for the restriction site, and clone it into the PUC57 vector to obtain the recombinant vector PUC57-SP-S-RBD-CTD.

Embodiment 3

[0067] The preparation method of the vector expressing novel coronavirus SARS-CoV-2 recombinant protein, comprises the following steps:

[0068] 1) Gene optimization synthesis

[0069] Link the N-terminal signal peptide SP, 306-575 region (S-RBD) of the new coronavirus SARS-CoV-2 S protein with the CTD region (aa248-365) of the N protein (S-RBD and CTD are connected with Linker GSGSG), According to the preference of the host cell Spodopterafrugiperda frugiperda (Sf9) cell line, the codon source was optimized, and PUC57 was used as the vector to Xho I , Kpn I Synthesize SP-S-RBD-CTD for the restriction site, and clone it into the PUC57 vector to obtain the recombinant vector PUC57-SP-S-RBD-CTD.

[0070] 2) Shuttle carrier preparation

[0071] Using Xho I, Kpn I, the SP-S-RBD-CTD in the vector PUC57-SP-S-RBD-CTD was digested and connected to the pFastBac TM P of the SCUVI plasmid polh under the promoter. Obtain the shuttle vector pFastBac TM SCUVI-SP-S-RBD-CTD.

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Abstract

The invention discloses a stable coronavirus recombinant protein dimer and its expression vector. The coronavirus recombinant protein consists of the coronavirus S protein S-RBD, the CTD region N-CTD of the coronavirus N protein and the connection connecting the two Sub composition. The coronavirus recombinant protein of some examples of the present invention can form and maintain a stable dimer structure, avoid the degradation of the monomer S-RBD, help improve the immunogenicity of the coronavirus recombinant protein, and is expected to be used for the preparation of raw materials for detection reagents and vaccines , antibodies, prophylactic or therapeutic drugs. The coronavirus recombinant protein dimers of some examples of the present invention have good immunogenicity. It has broad application prospects in the field of vaccine development. The expression vectors of some examples of the present invention are easy to express coronavirus recombinant protein dimers and have high expression levels.

Description

technical field [0001] The invention relates to a recombinant protein, in particular to a stable coronavirus recombinant protein dimer and its expression vector. Background technique [0002] Coronaviruses belong to the order Nidovirales, the family Coronaviridae, and the genus Coronavirus. They are important pathogens of many domestic animals, pets, and human diseases, causing a variety of acute and chronic diseases. According to the phylogenetic tree, coronaviruses can be divided into four genera: α, β, γ, and δ, among which β coronaviruses can be divided into four independent subgroups A, B, C and D. [0003] Coronavirus infections are extremely common around the world. So far, about 15 different coronavirus strains have been discovered, which can infect a variety of mammals and birds, and some can cause disease in humans. In addition to the recent outbreak of the new coronavirus SARS-CoV-2, there are six coronaviruses that can infect humans, including HCoV-229E, HCoV-O...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/165C12N15/50C12N15/63A61K39/215A61P31/14G01N33/569
CPCA61K39/12A61P31/14C07K14/005C12N2770/20022C12N2770/20034G01N33/56983G01N2333/165
Inventor 彭涛许煜华冯金郭伟丽陈丽云郑燕萍王炳彰刘双标
Owner SOUTH CHINA UNITED VACCINE INST
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