Preparation method of human lysozyme-antibacterial peptide Catesbeianin-1 fusion protein and application of same on preventing and curing cow mastitis
A technology of human lysozyme and fusion protein, applied in peptide/protein components, enzyme stabilization, drug combination, etc., can solve the problems of no patent disclosure, achieve low drug resistance, high activity, and enhance antibacterial and antiviral activity Effect
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[0062] Preparation of rhlyz-Catesbeianin-1 fusion protein
[0063] 1. Primer design and synthesis:
[0064] According to the human lysozyme gene sequence (NC-000012.10) and the antimicrobial peptide gene sequence published on Genebank, primers were designed: P1, P2, C1, C2, T1 and T2, and 15 flexible amino acid Linkers were designed to link the human lysozyme gene and the antimicrobial peptide Catesbeianin -1 gene concatenation. All primers were synthesized by Shanghai Bioengineering Company, and the primer sequences are as follows:
[0065] Human lysozyme upstream primer P1.5′-CG GAATTC ATGAAGGTCTTTGAAAGGTGTG-3' (introduced EcoR I restriction site)
[0066] Human lysozyme downstream primer P2: 5′-TT GCGGCCGC CACTCCACAACCTTGAACA-3′(Introduce Not I restriction site)
[0067] Catesbeianin-1 upstream primer C1: 5′-TCGGAAAAGGAGAGAAG-3′
[0068] Catesbeianin-1 downstream primer C2: 5′-GTAGGAATGTCACAGTAACG-3′
[0069] Specific upstream primer T1: 5′-GAAGGTCTTTGAAAGGTG-3′
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