Antibiotic resistance, especially in bacteria has motivated scientists to find novel bioactive compounds which may be used as antibiotic alternative. One of the most promising agents is antimicrobial peptide (AMP) which is a part of innate immune response. AMPs can be found virtually in all organisms but there are abundant in amphibians especially in frogs. This work focused on cloning AMP genes from Rana lateralis in order to get the sequence of putative peptides which were used for peptide synthesis. To assess therapeutic potential, each synthetic peptide was then used for antimicrobial peptide assay and hemolytic assay. The result indicated that 18-residue peptide RQ1 (ILPLLAGLVHGLSSIFGK) which has +2 net charge may be classified into Temporin family. This peptide can inhibit gram positive bacteria better than gram negative bacteria (MIC for Staphylococcus aureus and Bacillus cereus were 25 µM and 50 µM respectively). However, toxicity toward red blood cell was relatively high (HC50 = 60 µM). Thus, change of some amino acid sequence through peptide engineering is needed.