A foot-and-mouth disease virus non-structural protein 3b dominant epitope deletion-marked strain and its preparation method and application
A non-structural protein and foot-and-mouth disease virus technology, which is applied to the field of labeling strains with deletion of the dominant epitope of the non-structural protein 3B of foot-and-mouth disease virus and its preparation, can solve the problem of attenuated vaccine virulence regression, affecting vaccine immunogenicity, and large gene fragments. Problems such as virus replication ability
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[0051] 1. The construction method of the key amino acid deletion or modified full-length clone containing the dominant epitope of FMDV 3B protein:
[0052] The results of alanine scanning showed that the FMDV-specific 3B monoclonal antibody 4B1 recognized the key amino acid peptides of FMDV 3B1, 3B2 and 3B3 proteins as 2 PYAGP 6 . Using the FMD vaccine strain O / ZK / 93-08 infectious clone pOZKF-Z1234 as the backbone, using gene synthesis technology to mutate and modify the amino acids of the dominant epitopes of FMDV non-structural proteins 3B1, 3B2 and 3B3, and try to construct a 3B protein dominant strain FMD-tagged virus with complete depletion of the epitope.
[0053] In Jinweizhi Biological Co., Ltd. gene synthesis containing 3B protein amino acid mutation modification (see figure 1 ) OZK / 93-08 virus Z4 fragment, and cloned into pUC18 vector respectively, and the recombinant plasmids were respectively named pUC-Z43B6. The pUC-Z43B6 recombinant plasmid was digested with ...
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