Defensins are among the most important cationic peptides that their presence in a wide range of herbs and animals has been considered by their evolutionary role. these peptides have a prominent role in the inherent immune system. bean daphnia is one of these peptides, and on the other hand, given the importance of the development and introduction of new compounds inhibiting pathogenicity from nature, hence, in the present study, the recombinant synthesis of this peptide is secretion of the human embryonic kidney (hek293) cell line and its antimicrobial performance evaluation and its favorable results were obtained.
Methods
At first, codon optimization and subsequent adding of appropriate restriction sites to the vectors and codons. initiating and adding six coding sequences of the histidine and suitable end codon, we ordered the synthesis of the nucleotide sequences. in the next step, first, cutting the vector carrying the fragment with the enzyme and extracting the component from the gel and then cutting the vector with the same enzymes and the reaction of ligation and recombinant vector were created and after subclone in e.coli and extraction of plasmid, and then by culture of hek293 cells and transcript of recombinant pcdna3.1 + expression vector for hek293 and after antibiotic therapy and assembling the culture medium first confirmed the expression and secretion on the sds gel and finally the mic test.
Results
Confirmation of the ligation reaction with pcr colony was performed with the corresponding vector primers, and confirmation of the expression and secretion by sds gel and antimicrobial performance was also demonstrated with mic results in micrograms.
Conclusion
A clear perspective on the use of this peptide in inhibiting methicillin-resistant, staphylococcus aureus (mrsa) atcc 33591 and staphylococcus. aureus atcc 25923 and salmonella typhi and enterococcus faecalis atcc 29212
