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A kind of synthetic ebv consensus sequence molecular dna vaccine and its application

A DNA vaccine and sequence technology, applied in the field of genetic engineering technology and immunology, can solve the problems of limited efficiency and immaturity of chemical preparation liposome introduction

Active Publication Date: 2016-03-09
杭州中晟纳博生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The delivery efficiency of chemical formulations into liposomes is limited, and ultrasound and laser methods are still immature

Method used

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  • A kind of synthetic ebv consensus sequence molecular dna vaccine and its application
  • A kind of synthetic ebv consensus sequence molecular dna vaccine and its application
  • A kind of synthetic ebv consensus sequence molecular dna vaccine and its application

Examples

Experimental program
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Effect test

example 1

[0021] Example 1: Construction and expression identification of consensus sequence cEBNA1ΔGGA&GR expression vector.

[0022] Based on the sequences of EBNA1 proteins of different EBV strains, the consensus sequences were found. The EBNA1 sequences were derived from EBV strains B95-8, AG876 and GD1. The GGA and GR regions were removed from the EBNA1 consensus sequence, namely cEBNA1ΔGGA&GR, and its protein sequence was as follows:

[0023] cEBNA1ΔGGA&GR) amino acid sequence (372AA) encoded by SEQIDNo.1

[0024]MSDEGPGTGPGNGLGQKEDTSGPEGSGGSGPQRRGGDNHGRGRGRGRGRGGGRPGAPGGSGSGPRHRDGVRRPQKRPSCIGCKGAHGGTERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPPPGRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGRRKKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTEEGNWVAGVFVYGGSKTSLYNLRRGIALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLKDAIKDLVMTKPAPTCNIKVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDEGEEGQE

[0025] The N-terminal of cEBNA1ΔGGA&GR is connected with the leader sequence of IgE, and the C-termi...

example 2

[0026] Example 2: Immune responses produced by animals immunized with three kinds of plasmids pcEBNA1ΔGGA&GR.

[0027] Balb / c mice (six weeks old) were randomly divided into three groups (n=10). Two groups of animals were sedated by inhalation of isoflurane, the dorsal and ventral hairs were shaved, and DNA vaccine (pcEBNA1ΔGGA&GR10ug in 40ul1xPBS) was injected intradermally. One group received no other treatment after injection; The site of DNA injection was treated with electric pulse, 100V / cm, 2 pulse waves (BTXECM830, Harvard Apparatus, USA). After the initial immunization, booster immunization was given twice, with an interval of 3 weeks between each immunization. Blood samples were taken before each immunization, and serum was separated for ELISA detection of antibody immune response. One week after the last immunization, the mice were killed, and spleen mononuclear cells were collected from the spleen to detect the cellular immune response by ELISpot.

[0028] Anti-E...

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Abstract

The invention relates to three synthesized EBV (Epstein-Barr Virus) DNA (Deoxyribonucleic Acid) sequences which respectively encodes three consensus amino acid sequences EBNA1 (Epstein-Barr Nuclear Antigen 1), LMP1 (Latent Membrane Protein 1) and LMP2 (Latent Membrane Protein 2). The three sequences are respectively constructed on expression vector plasmids to express corresponding amino acid sequences. The invention further provides different methods for generating anti-corresponding EBV protein antigen immunoreaction by one or more expression plasmids.

Description

technical field [0001] The invention relates to the fields of genetic engineering technology and immunology technology, in particular to the consensus DNA sequence cloning, codon optimization, and eukaryotic vector expression construction of EBV virus-encoded protein EBNA1. The present invention is a therapeutic and preventive DNA vaccine product, that is, a synthetic Epstein-Barr virus (EBV) consensus (concensus) EBNA1 DNA sequence, which is constructed on an expression vector and applied to immunize the human body to induce the body's immune response , Generate cellular and humoral immunity against EBV virus infection or clear infected cells or cancerous cells containing EBV virus proteins. Background technique [0002] Epstein-Barr virus (EBV) causes more than 90% of infectious mononucleosis (IM) in developed countries. EBV is usually transmitted through oral secretions and infects B cells in the throat. The initial stage of infection is usually asymptomatic or non-speci...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/38C12N15/85A61K48/00A61P31/22
Inventor 林峰吴炯
Owner 杭州中晟纳博生物科技有限公司
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