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Construction of improved synergetic nucleic acid vaccine for preventing cow mastitis

A technology for nucleic acid vaccines and mastitis, applied in the field of improvement and construction of DNA vaccines, can solve the problems of unsatisfactory effect and low immunogenicity of nucleic acid vaccines, and achieve long duration of immune response, less dosage and enhanced effect Effect

Active Publication Date: 2013-01-02
XINJIANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, the effect of nucleic acid vaccines currently developed with FnBPA as the target gene is not very satisfactory. As a representative of a new generation of vaccines, low immunogenicity is a major factor limiting the widespread application of DNA vaccines

Method used

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  • Construction of improved synergetic nucleic acid vaccine for preventing cow mastitis
  • Construction of improved synergetic nucleic acid vaccine for preventing cow mastitis
  • Construction of improved synergetic nucleic acid vaccine for preventing cow mastitis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0033] Construction of eukaryotic expression vector of cow mastitis FnBPA gene

[0034] (1) Obtain the genomic DNA of the local epidemic strains.

[0035] (2) Screen the dominant epitopes of the FnBPA gene with good immunogenicity and antigenic diversity, and PCR amplify some genes of the A region and B region of the FnBPA gene (Figure 1), as the candidate sequence of the DNA vaccine.

[0036] (3) In order to add a signal peptide sequence to the amino terminal of Fn, synthesize primers according to the cow IFN-γ signal peptide cDNA sequence, FnP1-5'TGC TCT GTG GGC TTT TGG GTT TTT CTG GTT CTT ATG GCA CTA ACGTTA ATC ATA TAG 3' FnP2-5'CGA gga tcc ATG AAA TAT ACA AGC TAT TTC TTAGCT TTA CTG CTC TGT GGG CTT TTG GGT TTT TC 3'FnR5'-GGA Tga att cTTCAA TGT ATC CGT CAA C 3' and introduce BamHI in the upstream primer, downstream primer Two restriction sites of EcoR I were introduced, and the cow IFN-γ signal peptide sequence was fused with the FnBPA epitope gene by extension PCR.

[003...

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Abstract

The invention relates to a vaccine and a construction method thereof in the field of biotechnology, and is design, a construction method and an application of an improved synergetic nucleic acid vaccine. According to the invention, a fusion gene obtained by connecting an antigen epitope gene of staphylococcus aureus FnBPA with a GPI anchor sequence is inserted into a vector for expression and immunization; experiments show that the improved vaccine has good immunogenicity, and has better immune effect than single immunization of non-fusion genes. In attacking protection tests, immunoprotection effect is provided for immunized mice. The nucleic acid vaccine can prevent and control cow mastitis safely, effectively and with no medicament residues, and has good application prospects for the prevention and control of cow mastitis.

Description

technical field [0001] The invention relates to vaccines and construction methods in the field of biotechnology, in particular to the improvement and construction method of DNA vaccines targeting partial sequences of regions A and B of FnBPA molecules of endemic strains. Background technique [0002] In recent years, with the expansion of the breeding scale and the increase of the breeding density, the infection of mastitis is on the rise. As of 2011, the number of dairy cows in my country was 13 million, and the annual economic loss caused by mastitis can reach 4 billion yuan. In recent years, the extensive use or even abuse of antibiotics has gradually expanded the antibacterial spectrum of Staphylococcus aureus, posing new challenges to the clinical treatment and prevention of Staphylococcus aureus mastitis. Vaccination is currently considered to be an effective way to prevent and control Staphylococcus aureus infection. So far, the development of Staphylococcus aureus ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K39/085C12N15/62A61P15/14A61P37/04
Inventor 苏艳王世民张宝江
Owner XINJIANG AGRI UNIV