Patent ID: 8809036
Filing Date: 2014-08-19
Classification: C07K,C12N

Abstract:
1. A method of producing a recombinant antimicrobial peptide Cecropin AD (CAD) comprising the following steps: a. synthesizing the nucleic acid encoding a small ubiquitin-related modifier (SUMO) protease to produce a first synthetic DNA fragment; b. synthesizing the nucleic acid encoding the fusion protein of a c. digesting the first synthetic DNA fragment from step a with the restriction enzymes EcoR I and BamHI, and digesting the second synthetic DNA fragment from step b with the restriction enzymes BamHI and SacI; d. subcloning the products from step c into vector pBluesriptII SK(+) in order to produce a D1778-1 plasmid; e. extracting the genome of f. sub-cloning the amplified promoter p43 into a T-vector for sequencing; g. introducing the promoter p43 from the T-vector into pGJ103 plasmid to produce pGJ284 plasmid; h. digesting the pGJ284 plasmid and the D1778-1 plasmid with restriction enzymes EcoRI and SacI, respectively; i. recovering the two restriction products from step h, and then ligating said two products with T4 DNA ligase in water-bath at 16° C. to obtain a recombinant pNF11 plasmid; j. transforming k. inducibly expressing said pNF11 which comprises the nucleic acid encoding the antimicrobial peptide CAD in the recombinant