Τίτλος:
Characterization of pLAC1, a cryptic plasmid isolated from Lactobacillus acidipiscis and comparative analysis with its related plasmids
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
The pLAC1 plasmid of Lactobacillus acidipiscis ACA-DC 1533, a strain isolated from traditional Kopanisti cheese, was characterised. Nucleotide sequence analysis revealed a circular molecule of 3478. bp with a G. +. C content of 37.2%. Ab initio annotation indicated four putative open reading frames (orfs). orf1 and orf4 were found to encode a replication initiation protein (Rep) and a mobilization protein (Mob), respectively. The deduced products of orf2 and orf3 revealed no significant homology to other known proteins. However, in silico examination of the plasmid sequence supported the existence of a novel operon that includes rep, orf2 and orf3 in pLAC1 and that this operon is highly conserved also in plasmids pLB925A02, pSMA23, pLC88 and pC7. RT-PCR experiments allowed us to verify that these three genes are co-transcribed as a single polycistronic mRNA species. Furthermore, phylogenetic analysis of pLAC1 Rep and Mob proteins demonstrated that they may have derived from different plasmid origins, suggesting that pLAC1 is a product of a modular evolution process. Comparative analysis of full length nucleotide sequences of pLAC1 and related Lactobacillus plasmids showed that pLAC1 shares a very similar replication backbone with pLB925A02, pSMA23 and pLC88. In contrast, mob of pLAC1 was almost identical with the respective gene of plasmids pLAB1000, pLB4 and pPB1. These findings lead to the conclusion that pLAC1 acquired mob probably via an ancestral recombination event. Our overall work highlights the importance of characterizing plasmids deriving from non-starter 'wild' isolates in order to better appreciate plasmid divergence and evolution of lactic acid bacteria. © 2010 Elsevier B.V.
Συγγραφείς:
Asteri, I.-A.
Papadimitriou, K.
Boutou, E.
Anastasiou, R.
Pot, B.
Vorgias, C.E.
Tsakalidou, E.
Περιοδικό:
International Journal of Food Microbiology
Λέξεις-κλειδιά:
biological factor; mobilization protein; plasmid DNA; replication initiation protein; unclassified drug; bacterial protein, article; bacterial growth; bacterial strain; bacterium isolation; cheese; comparative study; copy number variation; Lactobacillus; Lactobacillus acidipiscis; nonhuman; nucleotide sequence; open reading frame; operon; phylogeny; plasmid; plasmid plac1; real time polymerase chain reaction; replicon; reverse transcription polymerase chain reaction; amino acid sequence; chemistry; classification; conformation; DNA base composition; genetics; isolation and purification; Lactobacillus; microbiology; molecular genetics; nucleotide sequence; plasmid; sequence alignment, Amino Acid Sequence; Bacterial Proteins; Base Composition; Base Sequence; Cheese; Lactobacillus; Molecular Sequence Data; Nucleic Acid Conformation; Open Reading Frames; Operon; Phylogeny; Plasmids; Sequence Alignment, Lactobacillus; Lactobacillus acidipiscis, Amino Acid Sequence; Bacterial Proteins; Base Composition; Base Sequence; Cheese; Lactobacillus; Molecular Sequence Data; Nucleic Acid Conformation; Open Reading Frames; Operon; Phylogeny; Plasmids; Sequence Alignment
DOI:
10.1016/j.ijfoodmicro.2010.05.018
