Antimicrobial activity of certain species of Lactobacillus bacteria were known to produce a certain type of bacteriocin called Plantaricin (Pln). In this study, the PlnF (plnF) gene (666 bp) encoding plantaricin was synthetically constructed based on a sequence of predefined and valuable amino acid compositions and subcloned into pET-28 (+) expression vector. Thereafter transformation of recombinant vector into BL21 (DE3) competent E. coli was achieved and to verify the result of cloning, the PCR colony technique was conducted. The expression of the inserted gene was analyzed by the SDS-polyacrylamide gel electrophoresis method, which detected a band of approximately 30 KD molecular weight for interested protein as a natural bioactive bacteriocin.