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Cell surface expression vector of SARS virus antigen and microorganisms transformed thereby

A surface expression, microorganism technology, applied in the direction of microorganisms, viruses/phages, double-stranded DNA viruses, etc., can solve the problem that the surface anchoring sequence has not been developed.

Inactive Publication Date: 2006-07-05
BIOLEADERS CORP +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no surface anchoring sequence satisfying the above conditions has been developed

Method used

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  • Cell surface expression vector of SARS virus antigen and microorganisms transformed thereby
  • Cell surface expression vector of SARS virus antigen and microorganisms transformed thereby
  • Cell surface expression vector of SARS virus antigen and microorganisms transformed thereby

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1: the synthesis of the antigenic site gene of the spike protein of SARS coronavirus

[0048] The spike protein of SARS coronavirus is a glycoprotein consisting of 1256 amino acids. In other coronaviruses that have been extensively tested, the spike protein is mostly embedded in the envelope protein that covers the surface of the virus particle, with an exposed structure. Exposure sites and antigenic sites have been intensively studied as target antigens for vaccines that induce viral infection and prevent infection.

[0049] Therefore, in order to select the site that can show antigenicity from the 1256 amino acids of the spike protein of SARS coronavirus, the spike of porcine transmissible gastroenteritis (TGE) coronavirus has been studied and synthesized for other Protein comparative analysis and structural comparative analysis were carried out to select antigenic sites. Specifically, it is known that the antigenic sites of the spike protein of porcine t...

Embodiment 2

[0070] Embodiment 2: the synthesis of the antigenic site gene of the nucleocapsid protein of SARS coronavirus

[0071] The nucleocapsid protein of SARS coronavirus is a protein consisting of 422 amino acids. Nucleocapsid proteins of most other coronaviruses have been reported to serve as antigens. These antigenic sites have been intensively studied for use as targeting antigens for vaccines against coronavirus infection.

[0072] Therefore, by carrying out protein comparative analysis with the nucleocapsid protein of porcine transmissible gastroenteritis (TGE), select the site that can show antigenicity in the amino acid of the nucleocapsid protein of SARS coronavirus, and carry out synthesis.

[0073] Specifically, the nucleocapsid protein of porcine transmissible gastroenteritis virus and the nucleocapsid of SARS coronavirus were analyzed according to the hydrophilicity map of the Kyte-Doolittle method, the antigenicity index of the Jameson-wolf method, and the surface like...

Embodiment 3

[0089] Embodiment 3: Construction of surface expression vector pHCE2LB: pgsA-SARS SA and pHCE2LB: pgsA-SARS SC

[0090]Taking Gram-negative bacteria and Gram-positive bacteria as hosts, using pgsA in the outer cell membrane protein gene (pgsBCA) derived from Bacillus strains and involved in the synthesis of poly-γ-glutamic acid to construct surface-expressing SARS The surface expression vectors pHCE2LB: pgsA-SARS SA and pHCE2LB: pgsA-SARS SC of the antigenic sites of the spike protein of the coronavirus SARS SA and SC.

[0091] First, digest pHCE2LB: pgsA-HPVL1 (KCTC 10349BP) with BamHI and KpnI to introduce the antigenic sites SARS SA and SARS SC in the spike protein of SARS coronavirus into Gram-negative bacteria and Gram-positive bacteria. On the surface expression carrier expressing the L1 antigen of human papillomavirus in the host of bacteria (a kind of carrier, it contains high expression promoter HCE promoter, the gene that participates in the outer cell membrane prote...

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Abstract

The present invention relates to a surface expression vector of SARS coronavirus antigen containing a gene encoding an antigen of SARS inducing coronavirus and any one or two or more of genes pgsB, pgsC and pgsA encoding poly-gamma-glutamic acid synthase complex, a microorganism transformed by the surface expression vector, and a SARS vaccine comprising the microorganism. According to the present invention, it is possible to economically produce a vaccine for prevention and treatment of SARS using a recombinant strain expressing an SARS coronavirus antigen on their surface.

Description

technical field [0001] The present invention relates to a carrier for expressing SARS antigen on the surface of microorganisms, a microorganism transformed by the carrier, and a vaccine for preventing SARS. The vaccine includes transformed microorganisms or their extracts and purified substances. More specifically, the present invention relates to a surface expression vector comprising a gene encoding an antigenic protein of a coronavirus that induces SARS, and one or two or more encoding microorganism surface anchoring sequences (anchoring motifs) The pgsB, pgsC and pgsA genes of the poly-γ-glutamic acid synthetase complex also relate to microorganisms transformed by the vector and SARS vaccines comprising the transformed microorganisms as active ingredients. Background technique [0002] Severe Acute Respiratory Syndrome (SARS) is a new type of epidemic disease. Since its first outbreak in Guangdong Province, China in November 2002, it has spread all over the world, includ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/86A61K39/00C07K14/025C07K14/165C12N9/00C12N15/74
CPCC12N9/93C07K2319/00A61K2039/523C12N2710/20022C12N2770/20022C12N15/74A61K39/00C07K14/005A61P31/12Y02A50/30C12N15/86
Inventor 成文喜金哲仲郑昌敏洪承杓李宗洙催在哲金光黑田俊一夫夏玲
Owner BIOLEADERS CORP
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