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Genetic engineering polypeptide vaccine for Myanmar subtype-containing 3 subtypes in O-type foot and mouth disease virus, its preparation and application

A foot-and-mouth disease virus and polypeptide vaccine technology, which is applied in the field of foot-and-mouth disease polypeptide vaccines, can solve the problems of long-term application, and achieve the effects of good safety, extended half-life, and large molecular weight

Inactive Publication Date: 2014-05-28
吴晓琰 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Despite many reports, it is far from being applied

Method used

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  • Genetic engineering polypeptide vaccine for Myanmar subtype-containing 3 subtypes in O-type foot and mouth disease virus, its preparation and application
  • Genetic engineering polypeptide vaccine for Myanmar subtype-containing 3 subtypes in O-type foot and mouth disease virus, its preparation and application
  • Genetic engineering polypeptide vaccine for Myanmar subtype-containing 3 subtypes in O-type foot and mouth disease virus, its preparation and application

Examples

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Embodiment 1

[0055] On the one hand, the present invention provides a genetically engineered polypeptide vaccine containing three subtypes of Myanmar subtype in the novel O-type foot-and-mouth disease virus, which contains the polypeptide encoded by the nucleic acid sequence shown in SEQ ID. SEQ ID contains the DNA sequence of 15 amino acid fragments of VP4 T-cell helper synthesized by chemical synthesis, the DNA sequence of three subtypes of VP1 B cell determinant and the DNA sequence of VP1 universal sequence [VP4 T helper- VP1(Wuhan 2006, GenBank: DQ478936.1)-VP1(Mya / 7 / 2002, GenBank: DQ164928.1)-VP1(O / Mya / 13 / 89, GenBank: DQ164924.1)-VP1(CN: 201010199712.2) ], cloned, and its expression product can stimulate the production of antibodies of three subtypes of O-type foot-and-mouth disease virus containing Myanmar subtype. see figure 1 , figure 2 , image 3 and Figure 4 .

[0056] In order to improve the bioavailability and enhance the immune effect, the vaccine of the present invent...

Embodiment 2

[0075] 1: Construction of expression plasmid

[0076] The genetically engineered bacteria are Escherichia coli containing the genes of the genetically engineered polypeptide vaccines of the three subtypes of the O-type foot-and-mouth disease virus including the Burmese subtype.

[0077] To synthesize the carrier of genetically engineered bacteria, in this example, the DNA sequence of the 15-amino acid fragment encoding the T-cell helper of VP4, the DNA sequence of the 3 subtypes of VP1 B cell determinants and the VP1 universal sequence were synthesized according to the O-type foot-and-mouth disease virus strain. The DNA sequences of [VP4T helper-VP1 (Wuhan 2006, GenBank: DQ478936.1)-VP1 (Mya / 7 / 2002, GenBank: DQ164928.1)-VP1 (O / Mya / 13 / 89, GenBank: DQ164924. 1)-VP1 (CN: 201010199712.2)], consisting of 4 DNA fragments (fragment 1: VP4T helper-VP1 (Wuhan 2006, GenBank: DQ478936.1); fragment 2: VP1 (Mya / 7 / 2002, GenBank: DQ164928). 1); Fragment 3: VP1 (O / Mya / 13 / 89, GenBank: DQ16492...

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Abstract

The invention relates to a genetic engineering polypeptide vaccine for Myanmar subtype-containing 3 subtypes in an O-type foot and mouth disease virus, its preparation and application. According to the invention, 2 common Myanmar subtypes (Mya / 7 / 2002, GenBank: DQ164928.1 and O / Mya / 13 / 89, GenBank: DQ164924.1) and another common O-type foot and mouth disease subtype (Wuhan 2006, GenBank: DQ478936.1) in the O-type foot and mouth disease virus are selected. A DNA sequence of VP4 T-cell helper 15 amino acid fragments and DNA sequences of B cell determinants of 3 subtype VP1, as well as the DNA sequence of a universal sequence (CN: 201010199712.2) of the O-type foot and mouth disease virus are connected in series, and cloning is carried out without carrier protein for constructing 1 genetic engineering bacterium. And after high density fermentation, cell disruption, inclusion body renaturation, fusion protein separation and purification, as well as freeze-drying, an antigenic protein product can be obtained. The antigenic protein product and adjuvant homogenate can form the polypeptide vaccine, which contains polypeptides encoded by nucleotide sequences shown in SEQ ID and has the advantages of good safety and high potency. One-time immunization can cover the 3 common subtypes of the O-type foot and mouth disease virus, so that the polypeptide vaccine can be produced in large scale without pollution.

Description

technical field [0001] The invention relates to a novel foot-and-mouth disease polypeptide vaccine, in particular to a genetically engineered polypeptide vaccine containing three subtypes of the Myanmar subtype in O-type foot-and-mouth disease virus and its preparation method and application. Background technique [0002] Foot and mouth disease (FMD) is an acute, febrile and highly contagious infectious disease of cloven-hoofed animals (pigs, cattle, sheep and camels) in animal husbandry, causing huge economic losses in the countries and regions where the epidemic occurs. loss. [0003] Foot-and-mouth disease is a global problem, and the epidemic has affected Germany (1937-1938), Turkey (1964-1965), Great Britain (1967-1968, 2007, 2009), Austria (1973), France (1974), South Korea (2000, 2002, 2010, 2011), Japan (2000, 2010), North Korea (2011) and other countries caused huge economic losses. At present, most countries in the world belong to "foot-and-mouth disease-affected...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/135A61K48/00A61P31/14C12N15/62C12N15/70C12R1/19
Inventor 吴晓琰赵泓龚铁军
Owner 吴晓琰
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