Chemosynthetic HSV2 virus gD glycoprotein extracellular region gene segment as well as expression and application thereof

A technology of chemical synthesis and gene fragments, which is applied in the fields of genetic engineering technology, vaccines and diagnostic reagents, and can solve problems such as limited clinical effects

Inactive Publication Date: 2010-07-28
李越希
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, a variety of HSV vaccines have been developed, and two HSV glycoprotein vaccines have entered Phase III clinical trials, namely the vaccine developed by Chiron’s recombinant gD2 / gB2 glycoprotein and MF59 adjuvant and Glaxo Smith Kline (GSK) The vaccine developed by the company is composed of recombinant gD2 glycoprotein and another adjuvant (3-de-O-acylated monophosphate lipid A and alum). Both vaccines have a certain protective effect, but the clinical effect is limited

Method used

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  • Chemosynthetic HSV2 virus gD glycoprotein extracellular region gene segment as well as expression and application thereof
  • Chemosynthetic HSV2 virus gD glycoprotein extracellular region gene segment as well as expression and application thereof
  • Chemosynthetic HSV2 virus gD glycoprotein extracellular region gene segment as well as expression and application thereof

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Embodiment Construction

[0045] Detailed description of the embodiment of the present invention:

[0046] Epitope Analysis, Gene Synthesis and Vector Construction of HSV2 Virus gD Glycoprotein Epitope

[0047] Analyze the entire amino acid sequence of the gD glycoprotein of the HSV2 virus by computer, screen out strong antigenic epitopes in the gD glycoprotein of the HSV2 virus, and select codons favored by both prokaryotic and eukaryotic, and chemically synthesize the gD glycoprotein of the HSV2 virus containing strong antigens An entirely new gene fragment for the epitope. The gene fragment was cloned into the Hind III / BamH I site in the plasmid pCEP4 to construct the recombinant eukaryotic expression vector pCEP4-gD2.

[0048] Materials and Methods

[0049] 1. Strains and plasmids:

[0050] Escherichia coli DH5α and eukaryotic expression vector pCEP4 are preserved by our laboratory.

[0051] 2. Reagents and instruments:

[0052] PrimerSTAR HS DNA Polymerase, dNTP, Hind III, BamH I, T4 DNA Liga...

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Abstract

The invention discloses a chemosynthetic HSV2 virus gD glycoprotein extracellular region gene segment as well as expression and application thereof, which relates to the technical field of gene engineering and the fields of vaccines and diagnosing agents. In the invention, through computer analysis, an HSV2 virus gD glycoprotein extracellular region segment containing strong antigen epitope is screened, and the HSV2 virus gD glycoprotein extracellular region segment contains 286 amino acid from the 1st amino acid to the 286th amino acid, and codons favored by both eucaryon and procaryote are selected to chemically synthesize a new gene sequence with the strong antigen epitope. Therefore, the gene segment is expressed by using the gene engineering technology, and the strong antigen epitope segment of HSV2 virus gD glycoprotein and antiserum of the HSV2 virus gD glycoprotein are prepared. The expressed HSV2 virus gD glycoprotein can be used for detecting vaccines, HSV2 virus antibody or antigen and preparing anti-HSV2 virus mono-antibody, poly-antibody and the like for immunity.

Description

technical field [0001] The chemically synthesized HSV2 virus gD glycoprotein extracellular region gene fragment and its expression and application in the present invention relate to the chemically synthesized HSV2 virus gD glycoprotein extracellular region gene fragment, using genetic engineering technology to prepare recombinant HSV2 virus gD sugar protein. Through computer analysis, the gD glycoprotein extracellular region fragment of HSV2 virus containing strong antigenic epitopes is screened out, the codons favored by both eukaryotic and prokaryotic organisms are selected, and a new gene sequence is chemically synthesized, which is expressed by genetic engineering technology. The protein can be used for the detection of vaccines and HSV2 virus antibodies or antigens, etc. The invention relates to the fields of genetic engineering technology, vaccines and diagnostic reagents. Background technique [0002] Herpes simplex virus (HSV) is the earliest discovered human herpes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/38C07K14/035A61K39/245A61P31/22G01N33/53C12R1/93
Inventor 李越希王正茂王忠灿管文燕李琳李素芹潘英潘明洁
Owner 李越希
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