Identification and cloning of a plasmid-encoded erythromycin resistance determinant from Lactobacillus reuteri

Axelsson, Lars; Ahrné, Siv; Andersson, Marie C.; Ståhl, Sten

doi:10.1016/0147-619x(88)90023-6

Cited by 67 publications

(30 citation statements)

References 18 publications

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“…Within the set of Tc r Lactobacillus isolates, the tet(M) genes of sequence homology group I and isolate DG 13 were found exclusively on plasmids (n ϭ 10), whereas for sequence homology group II, tet(M) genes were localized on the chromosome (n ϭ 4) or on a plasmid (n ϭ 10). R-plasmids encoding tetracycline, chloramphenicol, gentamicin, and macrolidelincosamide-streptogramin B resistance have been reported previously in L. reuteri (4,21,33,36), L. fermentum (10,18), L. acidophilus (36), and L. plantarum (1,8) isolated from raw meat, feces, and maize silage. Most of these R-plasmids were smaller than 10 kb.…”

Section: Discussionmentioning

confidence: 99%

Molecular Characterization of tet (M) Genes in Lactobacillus Isolates from Different Types of Fermented Dry Sausage

Gevers

Danielsen

Huys

et al. 2003

Appl Environ Microbiol

248

138

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The likelihood that products prepared from raw meat and milk may act as vehicles for antibiotic-resistant bacteria is currently of great concern in food safety issues. In this study, a collection of 94 tetracycline-resistant (Tc r ) lactic acid bacteria recovered from nine different fermented dry sausage types were subjected to a polyphasic molecular study with the aim of characterizing the host organisms and the tet genes, conferring tetracycline resistance, that they carry. With the (GTG) 5 -PCR DNA fingerprinting technique, the Tc r lactic acid bacterial isolates were identified as Lactobacillus plantarum, L. sakei subsp. carnosus, L. sakei subsp. sakei, L. curvatus, and L. alimentarius and typed to the intraspecies level. For a selection of 24 Tc r lactic acid bacterial isolates displaying unique (GTG) 5 -PCR fingerprints, tet genes were determined by means of PCR, and only tet(M) was detected. Restriction enzyme analysis with AccI and ScaI revealed two different tet(M) allele types. This grouping was confirmed by partial sequencing of the tet(M) open reading frame, which indicated that the two allele types displayed high sequence similarities (>99.6%) with tet(M) genes previously reported in Staphylococcus aureus MRSA 101 and in Neisseria meningitidis, respectively. Southern hybridization with plasmid profiles revealed that the isolates contained tet(M)-carrying plasmids. In addition to the tet(M) gene, one isolate also contained an erm(B) gene on a different plasmid from the one encoding the tetracycline resistance. Furthermore, it was also shown by PCR that the tet(M) genes were not located on transposons of the Tn916/Tn1545 family. To our knowledge, this is the first detailed molecular study demonstrating that taxonomically and genotypically diverse Lactobacillus strains from different types of fermented meat products can be a host for plasmid-borne tet genes.

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Section: Discussionmentioning

confidence: 99%

Molecular Characterization of tet (M) Genes in Lactobacillus Isolates from Different Types of Fermented Dry Sausage

Gevers

Danielsen

Huys

et al. 2003

Appl Environ Microbiol

248

138

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“…Micrococcus luteus A1 NCIMB 86166 (National Collection of Industrial and Marine Bacteria) was used as a nisin-sensitive indicator strain in nisin bioassays. Plasmid pLEB22 consisted of pUC6S (Viera & Messing, 1991) with an erythromycinresistance gene, erm (Axelsson et al, 1988), functional in L. lactis. Plasmid pKTH1980 (Graeffe et al, 1991) served as template for amplifying the nisZ gene.…”

Section: Bacterial Strains Plasmids Media and Growth Conditionsmentioning

confidence: 99%

NisB is required for the dehydration and NisC for the lanthionine formation in the post-translational modification of nisin

et al. 2002

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Nisin produced by Lactococcus lactis subsp. lactis is a 34-residue antibacterial polypeptide and belongs to a group of post-translationally modified peptides, lantibiotics, with dehydrated residues and cyclic amino acids, lanthionines. These modifications are supposed to be made by enzymes encoded by lanB and lanC genes, found only in biosynthetic operons encoding lantibiotics. To analyse the extent of modification, His-tagged nisin precursors were expressed in nisB and nisC mutant strains. The His-tagged nisin precursors were purified from the cytoplasm of the cells, as lack of NisB or NisC activity impaired translocation of the nisin precursor. The purified His-tagged polypeptides were analysed with trypsin digestion followed by nisin bioassay, SDS-PAGE, Nterminal sequencing and mass spectroscopy. According to the results, nisin precursors from the strain lacking NisB activity were totally unmodified, whereas nisin precursors from the strain lacking NisC activity, but having NisB activity, were dehydrated and devoid of normal lanthionine formation. This is the first experimental evidence showing that NisB is required for dehydration and NisC for correct lanthionine formation in nisin maturation.

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“…This vector is composed of the erythromycin resistance gene of Lb. reuteri (Axelsson et al, 1988), replication determinants obtained from the cryptic Lb. plantarum plasmid p256 (Cosby et al, 1989) and the pUC19 origin of replication.…”

Section: Introductionmentioning

confidence: 99%

Purification and amino acid sequence of sakacin A, a bacteriocin from Lactobacillus sake Lb706

Holck¹,

Axelsson²,

Birkeland

et al. 1992

Journal of General Microbiology

195

134

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Sakacin A, a bacteriocin produced by Lactobacillus sake Lb706 and which inhibits the growth of Listeria monocytogenes, was purified to homogeneity by ammonium sulphate precipitation and ion-exchange, hydrophobicinteraction and reversed-phase chromatography. The complete amino acid sequence of sakacin A was determined by Edman degradation. The bacteriocin consisted of 41 amino acid residues and had a calculated Mc of 4308.7, which is in good agreement with the value determined by mass spectrometry. The structural gene encoding sakacin A (sakA) was cloned and sequenced. The gene encoded a primary translation product of 59 amino acid residues which was cleaved between amino acids 18 and 19 to yield the active sakacin A. Sakacin A shared some sequence similarities with other bacteriocins.

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Identification and cloning of a plasmid-encoded erythromycin resistance determinant from Lactobacillus reuteri

Cited by 67 publications

References 18 publications

Molecular Characterization of tet (M) Genes in Lactobacillus Isolates from Different Types of Fermented Dry Sausage

Molecular Characterization of tet (M) Genes in Lactobacillus Isolates from Different Types of Fermented Dry Sausage

NisB is required for the dehydration and NisC for the lanthionine formation in the post-translational modification of nisin

Purification and amino acid sequence of sakacin A, a bacteriocin from Lactobacillus sake Lb706

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