Compound type therapeutic hepatitis B gene vaccine and construction method thereof

A technology of genetic vaccine and construction method, applied in the field of hepatitis B vaccine, can solve the problem of not being able to completely remove intracellular infection virus, etc., and achieve the effect of good intracellular virus and removal of intracellular virus

Inactive Publication Date: 2013-03-06
林绍强 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Preventive vaccines mainly induce the body to produce a humoral immune response. However, studies on viral diseases have found that humoral immunity cannot completely eliminate intracellular infected viruses, and only cell-mediated immune responses, especially cytotoxic T lymphocytes (CTL) mediated The cellular immune response can play the most fundamental role

Method used

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  • Compound type therapeutic hepatitis B gene vaccine and construction method thereof
  • Compound type therapeutic hepatitis B gene vaccine and construction method thereof
  • Compound type therapeutic hepatitis B gene vaccine and construction method thereof

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Experimental program
Comparison scheme
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Embodiment Construction

[0025] 1. PCR primer design and synthesis

[0026] Two pairs of primers were designed as the primer sequences for amplifying T-bet and HBV-L respectively. See Table 1 for the primer sequences and restriction sites. The sizes of the amplified T-bet and HBV-L gene fragments were 1608bp and 1203bp, respectively. Primers were synthesized by Invitrogen Company.

[0027] Target gene amplification primer sequence

[0028]

[0029] 2. PCR amplification of the target gene

[0030] 2.1 RT-PCR method to clone human T-bet gene

[0031] 2.1.1 Extraction of total RNA from human peripheral blood lymphocytes

[0032] (1) Take 5ml of anticoagulated blood from a healthy person with a blood collection tube, centrifuge at 3000rpm for 5min, and suck out the serum.

[0033] (2) Take a 15ml centrifuge tube and add 2ml Ficoll.

[0034] (3) Add 1ml PBS to the blood collection tube to resuspend the cells, attach to the wall at 45 degrees and add to a centrifuge tube equipped with Ficoll, and c...

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Abstract

The invention discloses a compound type therapeutic hepatitis B gene vaccine and a construction method thereof. The DNA vaccine contains a pIRES plasmid, a T-bet gene and a HBV-L gene, wherein the T-bet gene and the HBV-L gene are respectively located at multiple cloning sites (MCS) at two sides of an IRES element of the pIRES plasmid, the T-bet gene is located at MCSA, the HBV-L gene is located at MCSB, and nucleotide sequences of the T-bet gene and the HBV-L gene are respectively represented by SEQ ID No. 1 and SEQ ID No. 2. The DNA vaccine can be used for preventing and treating viral hepatitis type b.

Description

Technical field: [0001] The invention relates to a hepatitis B vaccine, specifically a composite therapeutic hepatitis B gene vaccine, and also relates to a construction method of the vaccine. Background technique: [0002] According to statistics, more than 2 billion people in the world have experienced HBV infection. Vaccination is the most effective measure to control and prevent hepatitis B and its long-term serious sequelae. Since the World Health Organization (WHO) recommended the implementation of hepatitis B vaccine program immunization in 1997, extensive vaccination has effectively reduced the rate of HBV infection. Cirrhosis, liver failure, and primary hepatocellular carcinoma due to chronic HBV infection. Prevention and treatment of HBV infection is imminent. [0003] At present, there are two intractable problems in the treatment of patients with hepatitis B infection: ①HBV exists in the liver for a long time and replicates to varying degrees, and the existing...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K39/29A61K39/39A61P31/20A61P1/16C12N15/12C12N15/51C12N15/63
Inventor 林绍强晏永波李晓霞刘乃华成军
Owner 林绍强
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