Preparation method of EV71 vaccine and vaccine prepared by preparation method
A technology for EV71 and vaccines, applied in the field of preparation of virus-like particle vaccines, which can solve the problems of long production cycle, high cost and low expression level
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[0031] 1. Gene selection and codon optimization design
[0032] Referring to the C4 type Chinese strain BJ08-Z020-1 (GenBank: FJ606449.1), DNA sequences encoding EV71P1 and 3CD were synthesized by techniques known in the art. The DNA sequences of BJ08-Z020-1 wild-type P1 and 3CD are SEQ ID NO: 1 and SEQ 1D NO: 2, and the amino acid sequences encoded by them are SEQ ID NO: 3 and SEQ ID NO: 4. The DNA sequences of wild-type P1 and 3CD were modified, and codons with high frequency in Pichia pastoris were used as codons to obtain optimized sequences SEQ ID NO: 5 and SEQ ID NO: 6.
[0033] 2. Construction of P13CD-pPEXZ recombinant expression vector
[0034] Firstly, the P1 and 3CD genes are obtained by means of gene synthesis known in the art according to the above synthesis method, and then the P1-pPICZaB and 3CD-pPICZaB recombinant vectors are constructed. Then, the AOX1 promoter of 3CD-pPICZaB was replaced with the pYPT1 promoter to obtain 3CD-pYPT1. The specific implementat...
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