SARS vaccine of adenovirus carrier and preparation method, application of coronavirus S gene

A coronavirus and vaccine preparation technology, applied in the field of biogenetic engineering, can solve problems such as infection and lack of vaccines

Inactive Publication Date: 2005-01-12
SUN YAT SEN UNIV CANCER CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] According to current research, among the four known structural proteins of coronaviruses, the spike protein (spike S) is a structural protein that induces a protective immune response. The research structure of some scholars has confirmed

Method used

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  • SARS vaccine of adenovirus carrier and preparation method, application of coronavirus S gene
  • SARS vaccine of adenovirus carrier and preparation method, application of coronavirus S gene
  • SARS vaccine of adenovirus carrier and preparation method, application of coronavirus S gene

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The preparation method of adenovirus vector SARS vaccine:

[0036] The preparation of the adenovirus vector SARS vaccine is divided into two parts: the early construction and the later amplification:

[0037] Early build:

[0038] Get the SARS-related coronavirus spike(S F , S N , S M , S C ) gene, amplified by PCR method, after PCR, digested with Xbal+Kpnl 37°C, and simultaneously digested pShuttle with this enzyme, ligated, transformed into Escherichia coli DH5a, and used kanamycin (Kan R ) resistance screening positive clones, cultured and purified to obtain pS F / S N / S M / S C -Shuttle, use 1-Ceul+P1-Scel to digest, and use this enzyme to digest PAdeno-X TM , ligated after digestion, transformed into Escherichia coli DH5a, using ampicillin (Amp + ) resistance screening positive clones to obtain pAd-SF / S N / S M / S C .

[0039] Expand training:

[0040] get pAd-S F / S N / S M / S C Afterwards, after digestion with Pacl and linearization of the plasmi...

Embodiment 2

[0049] Cloned into the adenovirus vector is the N-terminal fragment of the S gene of SARS-associated coronavirus. All the other are with embodiment 1.

Embodiment 3

[0051] Cloned into the adenovirus vector is the middle fragment of the SARS-associated coronavirus S gene. All the other are with embodiment 1.

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PUM

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Abstract

The invention pertains to biological genetic engineering field, specificly relating to SARS vaccine of adenovirus carrier and preparation method, application of related coronavirus S gene in preparation of the SARS vaccine for preventing SARS. Through a bioengineering means, the related coronary virus S gene and a defective adenovirus are recombined, which makes protective immunogen protein or polypeptide expressing in it. A genic vaccine that can arouse the mucosal immunogenicity is produced through amplifying training, purification, and preparation, which induces a immunity reaction in the respiratory mucosa, makes the organism produce the corresponding antibody, and prevents virus from infection. Compared with the traditional inactivated viral particles vaccine, the invention has a high safety; it is convenient to operate; it is not limited by the specific conditions such as intramuscular injection; and it has an extensive clinical application prospect.

Description

1. Technical field [0001] The invention belongs to the field of biogenetic engineering, and in particular relates to an adenovirus vector SARS vaccine, a preparation method thereof, and an application of revealing a related coronavirus S gene in preparing a vaccine for preventing atypical pneumonia (SARS). 2. Background technology [0002] The outbreak of atypical pneumonia (atypical pneumonia) began in the Guangdong area of ​​China at the end of last year. At present, the disease has spread to more than 30 countries and regions, and the WHO has named it severe acute respiratory syndrome (severe acute respiratory syndrome, SARS). [0003] At present, scientists from many countries in the world have independently isolated new coronaviruses (coronavirus) from the sera of different patients. The results of virus gene sequence analysis show that they have 50% to 60% homology with known coronaviruses. The World Health Organization (WHO) has announced that the pathogen causing SA...

Claims

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Application Information

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IPC IPC(8): A61K39/215A61K48/00A61P11/00A61P31/14C12N15/861
Inventor 黄文林曾益新汪健刘然义黄嘉凌黄必军赖坤吴立志梁志慧柯妙拉吴秀菊
Owner SUN YAT SEN UNIV CANCER CENT
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