A kind of o/asia type I foot-and-mouth disease virus bivalent genetic engineering polypeptide vaccine and its preparation method and application

A foot-and-mouth disease virus and genetic engineering technology, which is applied in the field of O/Asia Ⅰ type foot-and-mouth disease virus bivalent genetically engineered polypeptide vaccine and its preparation, can solve the problems of remote application and achieve the effects of good safety, high yield and low cost

Inactive Publication Date: 2011-12-14
吴晓琰 +2
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  • Abstract
  • Description
  • Claims
  • Application Information

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  • A kind of o/asia type I foot-and-mouth disease virus bivalent genetic engineering polypeptide vaccine and its preparation method and application
  • A kind of o/asia type I foot-and-mouth disease virus bivalent genetic engineering polypeptide vaccine and its preparation method and application
  • A kind of o/asia type I foot-and-mouth disease virus bivalent genetic engineering polypeptide vaccine and its preparation method and application

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Embodiment 1

[0035] According to the amino acid sequence of the VP1 immune determinant of O and Asia I two types of FMDV and the T-cell helper of VP4, the present invention designs and synthesizes DNA gene fragments. Its genetically engineered bacteria is O / AsiaⅠ. O / AsiaⅠgenetic engineered bacteria is the DNA sequence of 15 amino acid fragments of the T-cell helper encoded by the foot-and-mouth disease virus strain of VP4, which is connected to the immunogenic immunodeterminant region of the O-type VP1 protein. The DNA sequence is connected to the AsiaⅠtype VP1 protein. The DNA sequence of the immunogenic immunodeterminant region as 1 repeat, the nucleotide sequence of 1 repeat can be found in figure 1 .

[0036] The novel O / Asia I foot-and-mouth disease virus bivalent genetically engineered polypeptide vaccine of the present invention contains 2 n-1 A polypeptide encoded by the nucleic acid sequences shown in the concatenated SEQ ID, wherein n is an integer of 1-5. SEQ ID contains the ...

Embodiment 2

[0060] 1: Construction of expression plasmids

[0061] Genetically engineered bacteria are O / Asia I genetically engineered bacteria.

[0062] In order to synthesize the vector of O / Asia I genetically engineered bacteria, the DNA sequence of the 15 amino acid fragments of the VP4 T-cell helper encoded by the foot-and-mouth disease virus strain was connected with the DNA sequence of the immunogenic immunodeterminant region in the O-type VP1 protein The DNA sequence of the immunogenic immunodeterminant region in the Asia I type VP1 protein is regarded as a repeating fragment F1, which is produced by connecting 10 DNA fragments, and each repeating fragment F1 contains figure 1 Nucleic acid sequences and appropriate restriction enzyme cut sites are shown. Use genetic engineering to clone F1 into the Lac promoter expression plasmid, and then follow the figure 2 According to the method, F2, F4, and F8 can be cloned into Lac promoter expression plasmids sequentially, which contain ...

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Abstract

The invention relates to an O/Asia I type foot and mouth disease virus bivalent genetic engineering polypeptide vaccine, its preparation method and its purpose. The method comprises the following steps: selecting two serotypes of O type and Asia I type, taking B cell determinant 15 amino acid fragments of VP1 and T-cell helper of VP4, performing a series connection, cloning without containing carrier protein, constructing O/Asia I gene engineering bacteria. An antigen protein product can be obtained after passing through the processes of high density fermenting, cell disrupting, inclusion body renaturating, fusion protein separating, and is homogenized with an adjuvant to form the O/Asia I type foot and mouth disease virus bivalent genetic engineering polypeptide vaccine. The vaccine of the present invention contains 2<n-1> polypeptide connected in series which is coded by a nucleic acid sequence shown in SEQ ID, wherein, n is an integer of 1-5. The invention has the advantages of good security and high immune efficacy, and can be used once in half year for immunization; and is suitable for large scale production and convenient preservation and transportation; and is capable of effectively preventing and controlling two serotypes foot and mouth disease of O type and Asia I type which is useful in our country; foot and mouth disease virus non-structural protein 3A.B. will not generate, so that the infective animals can be differentiated easily.

Description

technical field [0001] The invention relates to a novel foot-and-mouth disease vaccine, in particular to a bivalent gene engineering polypeptide vaccine of type O / Asia I foot-and-mouth disease virus produced by a genetic engineering method and its preparation method and application. Background technique [0002] The continuous expansion of domestic breeding scale will become the source of continuous growth for the animal vaccine industry. At present, my country's meat and egg production ranks first in the world, and it is the world's largest producer of livestock products. In 2004, the annual output value of my country's animal husbandry exceeded 1 trillion yuan for the first time, and the proportion of the output value of animal husbandry to the total agricultural output value was gradually increasing. Due to the continuous epidemics worldwide, foot-and-mouth disease, bird flu, mad cow disease, etc. have brought challenges to the prevention and control of animal diseases i...

Claims

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Application Information

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IPC IPC(8): A61K39/135C12N1/21C12N15/62C12N15/70C07K19/00A61P31/14C12R1/19
CPCY02P20/582
Inventor 吴晓琰赵泓孙玉琨
Owner 吴晓琰
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