Chemically synthesized HSV1 virus gD glycoprotein extracellular region gene fragment and expression and application thereof

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

Inactive Publication Date: 2010-02-10
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

Method used

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  • Chemically synthesized HSV1 virus gD glycoprotein extracellular region gene fragment and expression and application thereof
  • Chemically synthesized HSV1 virus gD glycoprotein extracellular region gene fragment and expression and application thereof
  • Chemically synthesized HSV1 virus gD glycoprotein extracellular region gene fragment and expression and application thereof

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Experimental program
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Effect test

Embodiment Construction

[0106] Detailed description of the embodiment of the present invention:

[0107] Analysis, gene synthesis and vector construction of gD glycoprotein epitope of HSV1 virus

[0108] Analyze the entire amino acid sequence of the gD glycoprotein of the HSV1 virus by computer, screen out strong antigenic epitopes in the gD glycoprotein of the HSV1 virus, and select codons favored by both prokaryotic and eukaryotic, and chemically synthesize the gD glycoprotein of the HSV1 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-gD1.

[0109] Materials and Methods

[0110] 1. Strains and plasmids:

[0111] Escherichia coli DH5α and eukaryotic expression vector pCEP4 are preserved by our laboratory.

[0112] 2. Reagents and instruments:

[0113] PrimerSTAR HS DNA Polymerase, dNTP, Hind III, BamH I, T4 DNA Ligase, P...

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Abstract

The invention discloses a chemically synthesized HSV1 virus gD glycoprotein extracellular region gene fragment and expression and an application thereof, and relates to the fields of genetic engineering technology, vaccine and diagnostic reagent. By computer analysis, the invention screens out the HSV1 virus gD glycoprotein extracellular region fragment containing strong antigen epitope, totally 284 amino acids, namely from the 1st to the 284th; codons which are preferred by both eukaryon and pronucleus organism are selected; and a brand new gene sequence of the antigen epitope is chemically synthesized; the genetic engineering technology is utilized to express the gene fragment and prepare the strong antigen epitope fragment and antiserum of the HSV1 virus gD glycoprotein. The expressed HSV1 virus gD glycoprotein can be applicable to the detection of vaccine, HSV1 virus antibody or antigen and immune preparation of HSV1 virus monoclonal antibody and polyclonal antibody and the like.

Description

technical field [0001] The invention is a brand-new gene segment of the extracellular region of gD glycoprotein of HSV1 virus chemically synthesized, and uses genetic engineering technology to prepare recombinant HSV1 virus gD glycoprotein. Through computer analysis, the gD glycoprotein extracellular region fragment of HSV1 virus containing strong antigenic epitope is screened out, codons favored by both eukaryotic and prokaryotic organisms are selected, a new gene sequence is chemically synthesized, and expressed by genetic engineering technology. The protein can be used for the detection of vaccines and HSV1 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 virus, which can easily lead to recurrent infection and latent infection, and is seriously harmful to the human body. It is divided in...

Claims

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Application Information

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IPC IPC(8): C12N15/38C07K14/035A61K39/245A61P31/22G01N33/53
Inventor 李越希王正茂管文燕李琳李素芹潘英潘明洁
Owner 李越希
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