Biological preparation for resisting against replication and infection of foot-and-mouth disease virus and preparation method as well as application thereof
An anti-foot-and-mouth disease virus and biological agent technology, which is applied in the directions of biochemical equipment and methods, antiviral agents, and botanical equipment and methods, can solve the problems of difficulty in achieving the protective effect of mutant strains, and achieves inhibition of foot-and-mouth disease virus replication and inhibition. The effect of anti-foot-and-mouth disease infection
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[0060] The present invention is based on O-type FMDV isolated from different ages and regions, selects highly conserved sequences of FMDV 2B and VP1 genes, designs and synthesizes siRNA, clones them into the PSIREN-shuttle shuttle vector, and constructs recombinant plasmids pShuttle-2B and pShuttle-VP1. The specific process as follows:
[0061] (1) Dilute the transcription template of siRNA-VP1 and the transcription template of siRNA-2B to the same concentration (5pmol / μL) with deionized water, and mix the two complementary oligonucleotides (5μL each) Then put it on the PCR amplification instrument, treat at 95°C for 2min, and then cool down to normal temperature naturally for processing. The double-stranded siRNA oligonucleotide fragments formed after treatment were respectively ligated with pSIREN-Shuttle (Clontech Company) after double digestion with BamHI and EcoRI (the ligation reaction was operated according to the instructions of T4 DNA ligase, and the screening of posi...
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