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.

CN1488646AInactive Publication Date: 2004-04-14李越希
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2004-04-14
Estimated Expiration
Not applicable · inactive patent

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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.
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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...

Examples

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 ...