Konstruksi Gen Penyandi Plantaricin S dari Lactobacillus plantarum S34

Abstract

Bacteriocins are antibacterial peptides that commonly used as food biopreservative. Bacteriocins were produced by both either Gram positive or Gram negative bacteria, one of them is Lactobacillus plantarum S34 isolated from beef bekasam. L. plantarum S34 produced bacteriocins called plantaricin. PlnS gene is gene that encoding plantaricin. The study purposed is to isolate plnS gene from L. plantarum S34. The plnS gene amplified by PCR (Polymerase Chain Reaction) technique, and then cloned into cloning vector pGEM-T Easy and transformed into E. coli DH5α. Transformation gene using heat shock method. Selection of transformants using the blue and white colonies. Gene amplification produces plnS gene amplicon size about ±500 bp. PlnS gene ligation with pGEM-T Easy (3015 bp) vector generating plasmid size about ±3500 bp. The confirmation of white colonies from blue-white screening was carried out with PCR colony, generated amplicons size was about ±500 bp. Recombinant plasmid in colony 2 was confirmed by restricition analysis using SalI and NcoI. The result showed that the gene insert had expected size ±500 bp.