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SARS virus S protein and N protein fusion protein, and preparation and use thereof

A SARS virus, fusion protein technology, applied in antiviral immunoglobulins, DNA/RNA fragments, antiviral agents, etc., can solve problems such as difficulty in mass production, false positives, and no vaccine to prevent SARS.

Inactive Publication Date: 2004-04-14
李越希
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The SARS virus can be cultured in vitro with Vero-E6, so the immunofluorescence method and enzyme-linked immunoassay method for detecting SARS virus antibodies have been established using the cultivated SARS virus, but it is difficult to mass-produce the antigen detection antibody using the cultured SARS virus, and it will also False positives are generated, and the development of specific genetic recombinant antigens is the development direction of the establishment of SARS virus-specific antibody detection reagents
The most ideal way to prevent SARS infection is to inject vaccines, but there is no vaccine to prevent SARS at present, and the development of vaccines is being actively carried out at home and abroad, especially the research on inactivated vaccines is progressing rapidly, and genetic engineering vaccines are the ultimate development direction

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  • SARS virus S protein and N protein fusion protein, and preparation and use thereof
  • SARS virus S protein and N protein fusion protein, and preparation and use thereof
  • SARS virus S protein and N protein fusion protein, and preparation and use thereof

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Embodiment Construction

[0049] Detailed description of the embodiments of the present invention:

[0050] Analysis, gene synthesis and expression of S protein and N protein epitopes of SARS virus

[0051] Through computer analysis of the amino acid sequence of SARS virus N protein and S protein, the N protein fragments of SARS virus with strong epitopes were screened, namely the first amino acid to 258 amino acids, and the S protein of SARS virus with strong epitopes Fragment, namely the 162nd amino acid to the 460th amino acid, a total of 299 amino acids. Choose codons favored by both eukaryotes and prokaryotes, chemically synthesize brand-new gene sequences of SARS virus N protein fragment and S protein fragment, and use genetic engineering technology to connect the two gene fragments in series and clone into the NcoI in plasmid pET28a(+) / EcoRI site, expresses the fusion protein of S protein fragment and N protein fragment. The recombinant plasmid was transformed into Escherichia coli BL21 (DE3), and ...

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Abstract

The invention is confluens protein of SARS virus S protein and N protein, and the manufacturing, application and the gene project technology, vaccine and diagnosing reductant. Through analyzing the cistine series of SARS s protein and N protein, sifts out SARS virus S protein segment with strong antigen epitope, namely the 162nd cistine to 460nd cistine, and the N protein segment of SARS virus with strong antigen epitope, anmely the first cistine to 258nd cistine. It synthesizes the new SARS virus S protein segment and N protein segment gene series, connects the two gene segments in series. Expresses the confluens protein of the two protein segments, the two are connected by glycine and serine, inserts a glycine adjoining to the frist cistine methionine of N protein segment, the confluens proteiní» length is 560 cistines.

Description

Technical field [0001] The invention connects the chemically synthesized brand-new gene segment of the SARS virus S protein and the gene segment of the N protein, and uses genetic engineering technology to prepare the fusion protein of the SARS virus S protein and the N protein. Through computer analysis, the fusion protein and N protein fragments of the S and N protein fragments of SARS virus with strong epitopes were screened out, the codons favored by both eukaryotic and prokaryotes were selected, and a new S gene sequence and sequence were chemically synthesized. The N gene sequence uses genetic engineering technology to express the fusion protein of the two. The expressed fusion protein can be used for vaccines and the detection of SARS virus antibodies or antigens. The present invention relates to the fields of genetic engineering technology, vaccines and diagnostic reagents. Background technique [0002] Severe Acute Respiratory Syndrome, also known as SARS (Severe Acute R...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/215A61P11/00A61P31/14C07K16/08C07K19/00C12N15/62C12N15/63G01N33/68
Inventor 李越希陶开华朱敏生
Owner 李越希
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