BACKGROUND AND OBJECTIVES The Unlimited use of antibiotics nowadays has led to many problems, including the appearance of antibiotic-resistant bacteria. For this reason, the use of antimicrobial peptides can be a good alternative to antibiotics. The aim of this research is to design and manufacture a chimeric antimicrobial peptide and transfer it to a suitable vector. MATERIALS AND METHODS At first, a search was made in scientific databases to find antimicrobial peptides. Next, the appropriate peptide was selected using the docking process. The nucleotide sequence of the selected peptide was optimized for better performance and compatibility in the direction of production in E. coli. The selected nucleotide sequence was inserted into pET26 vector. After the plasmid containing this sequence was prepared, the stages of preparation and growth of the host bacteria were carried out. Then transformation processes were carried out into the bacterial host using heat shock. RESULTS AND DISCUSSION The growth of bacteria in the positive transfer samples and the lack of growth of bacteria in the negative transfer samples indicate that the plasmid was received by the expression vector, and the PCR process was also done to ensure that. The production of this recombinant peptide was also done under optimal induction conditions. CONCLUSION Optimizing the expression conditions for chimeric antimicrobial peptide production was done using pET26 vector by host Escherichia coli strain Bl21.which can be used for therapeutic purposes to combat antibiotic-resistant bacteria.