Skip to main content
Fig. 1 | BMC Microbiology

Fig. 1

From: Identification and characterization of the bacteriocin Carocin S3 from the multiple bacteriocin producing strain of Pectobacterium carotovorum subsp. carotovorum

Fig. 1

Flow diagram of the research process. To confirm the presence of another bacteriocin from Pcc, we initially subjected the TH22–10 (caroS1K:Tn5) to bacteriocin activity assay, RT-PCR, and western blotting. After confirming that the Pcc strain H-rif-8-6 produces another bacteriocin apart from the previously identified bacteriocin, Carocin S1, the genomic DNA from H-rif-8-6 was isolated and digested using several restriction enzymes and was assayed with the best restriction enzyme for creating DNA libraries as demonstrated by the Southern blotting experiment. Then, bacteriocin production was tested through the bacteriocin activity assay. After that, the DNA nucleotide sequence and the deduced amino acid sequence of other known bacteriocins were compared. Subsequently, subcloning for the novel bacteriocin, designated as Carocin S3, and transformation were performed. Recombinants were used to express the Carocin S3 proteins, Carocin S3K and Carocin S3I. Furthermore, bacteriocin expression and bacteriocin activity assays were performed. Also, protein purification and Carocin S3 antibiotic activity test were carried out. Finally, Electrospray Ionization Mass Spectrometry Molecular Weight Assay was done to determine the molecular weights of the killer protein, Carocin S3K, and immunity protein, Carocin S3I

Back to article page