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Preparation method of periodic wuchereria malayi compound multivalent DNA (deoxyribonucleic acid) vaccine

A DNA vaccine, Malayan filarial technology, applied in the field of periodic Malayan filarial vaccine preparation, can solve the problems of difficulty in inducing sufficient and effective immune protection, life history and complex antigen molecules

Inactive Publication Date: 2013-07-10
NANTONG UNIVERSITY
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  • Application Information

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Problems solved by technology

[0004] Filarial worms are multicellular human parasites with complex life history and antigenic molecules, and have developed a variety of immune evasion mechanisms during the long-term evolution with the host, so it is difficult for monovalent vaccines to successfully induce sufficient and effective immune protection in vivo

Method used

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  • Preparation method of periodic wuchereria malayi compound multivalent DNA (deoxyribonucleic acid) vaccine
  • Preparation method of periodic wuchereria malayi compound multivalent DNA (deoxyribonucleic acid) vaccine
  • Preparation method of periodic wuchereria malayi compound multivalent DNA (deoxyribonucleic acid) vaccine

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Embodiment Construction

[0136] 1 Primer design

[0137] According to the known sequence of BmGAPDH gene in GeneBank, primers were designed using Primer Premier5.0 software, and BamH Ⅰ and Xho Ⅰ were introduced at the 5′ ends of the upstream and downstream primers respectively. Restriction sites, start, stop codons and protective bases, primers were synthesized by Shanghai Sangon Bioengineering Company.

[0138] BmGAPDH-P1: upstream primer 5'-CC GGATCC ACC ATG ATT AAC ATT GAC TAT-3'underlined sequence is BamH Ⅰ restriction site (Tm=62.02)

[0139] BmGAPDH-P2: downstream primer 5'-CC CTCGAG TTA GGT TGC TGT AGC CAT AT-3'The underlined sequence is the XhoⅠ restriction site (Tm=64.94)

[0140] According to the known sequence of Cysteine ​​Protease Inhibitor (BmCPI) gene in GeneBank, primers were designed using Primer Premier5.0 software, and Nhe Ⅰ and BamH Ⅰ enzymes were introduced at the 5' ends of the upstream and downstream primers respectively. Cut site, start stop codon and protective bases, p...

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Abstract

The invention discloses a preparation method of a periodic wuchereria malayi compound multivalent DNA (deoxyribonucleic acid) vaccine. The preparation method comprises the following steps of: designing a primer, amplifying BmGAPDH (reduced glyceraldehyde-phosphate dehydrogenase) and BmCPI genes by PCR (polymerase chain reaction), purifying segments of BmGAPDH and BmCPI genes, connecting and converting a T carrier, screening and identifying an I positive recombinant clone, building and identifying an I recombinant expression vector, extracting and purifying a mass of recombinant plasmid and the like. The preparation method is convenient in preparation and good in effect. The periodic wuchereria malayi which is popular in China is taken as a research object, and the periodic wuchereria malayi cysteine proteinase inhibitor / 3-phosphoglyceraldehyde dehydrogenase eukaryotic expression recombinant plasmid is built. The test result proves that the compound multivalent DNA vaccine can induce the mouse to generate the specific immune response reaction, so that a satisfied effect can be obtained, and the periodic wuchereria malayi compound multivalent DNA vaccine can be successfully prepared.

Description

technical field [0001] The invention relates to a preparation method of a periodic type malayia filarial vaccine. Background technique [0002] Filariasis is a vector-borne disease that seriously endangers human health. It is jointly listed by the United Nations Development Program / World Bank / World Health Organization as the world's key prevention and control of malaria, schistosomiasis, leishmaniasis, trypanosomiasis, and leprosy. One of the six major tropical diseases outside the disease, lymphatic filariasis is prevalent in more than 70 countries and regions in the world, and 1.1 billion people are threatened by infection. Chronic filariasis mainly causes elephantiasis, hydrocele and chyluria, etc., which can cause patients to lose their ability to work to varying degrees. It is listed by the World Health Organization as the second disabling disease in the world, second only to mental disorders. There are more than 90 million lymphatic filariasis patients in the world, a...

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Application Information

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IPC IPC(8): A61K39/00C12N15/85C12N15/66A61P33/10
CPCY02A50/30
Inventor 方政王慧方浩陆施娟胡迎青徐邦生
Owner NANTONG UNIVERSITY
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