title: Expression of bacteriocin LsbB is dependent on a transcription terminator.
abstract: The production of LsbB, leaderless class II bacteriocin, is encoded by genes (lsbB and lmrB) located on plasmid pMN5 in Lactococcus lactis BGMN1-5. Heterologous expression of the lsbB gene using the pAZIL vector (pAZIL-lsbB) in L. lactis subsp. cremoris MG7284 resulted in a significant reduction (more than 30 times) of bacteriocin LsbB expression. Subcloning and deletion experiments with plasmid pMN5 revealed that full expression of LsbB requires the presence of a complete transcription terminator located downstream of the lsbB gene. RNA stability analysis revealed that the presence of a transcription terminator increased the RNA stability by three times and the expression of LsbB by 30 times. The study of the influence of transcription terminator on the expression of other bacteriocin genes (lcnB, for lactococcin B production) indicated that this translational terminator likely functions in a lsbB-specific manner rather than in a general manner.
authors.affiliation: Laboratory for Molecular Microbiology, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Serbia; Faculty of Agriculture, University of Belgrade, Serbia.
authors.forename: Gordana
authors.initials: G
authors.lastname: Uzelac
authors.affiliation: Laboratory for Molecular Microbiology, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Serbia.
authors.forename: Marija
authors.initials: M
authors.lastname: Miljkovic
authors.affiliation: Laboratory for Molecular Microbiology, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Serbia; Faculty of Biology, University of Belgrade, Serbia.
authors.forename: Jelena
authors.initials: J
authors.lastname: Lozo
authors.affiliation: Faculty of Agriculture, University of Belgrade, Serbia.
authors.forename: Zorica
authors.initials: Z
authors.lastname: Radulovic
authors.affiliation: Laboratory for Molecular Biomedicine, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Serbia.
authors.forename: Natasa
authors.initials: N
authors.lastname: Tosic
authors.affiliation: Laboratory for Molecular Microbiology, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Serbia. Electronic address: mkojic@imgge.bg.ac.rs.
authors.forename: Milan
authors.initials: M
authors.lastname: Kojic
mesh_terms: D001430:Bacteriocins / Q000096:biosynthesis* / Q000235:genetics; D004269:DNA, Bacterial; D015964:Gene Expression Regulation, Bacterial; D055786:Gene Knockout Techniques; D005798:Genes, Bacterial; D013294:Lactococcus lactis / Q000201:enzymology / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic
chemical_list: D001430:Bacteriocins; D004269:DNA, Bacterial; C075150:lactococcin B
keywords: Bacteriocin expression; LsbB baceriocin; Transcription terminator regulation