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

[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 ...

[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 ...