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. 2005 Mar;71(3):1223-30.
doi: 10.1128/AEM.71.3.1223-1230.2005.

Functional analysis of three plasmids from Lactobacillus plantarum

Richard van Kranenburg  1 , Natasa Golic, Roger Bongers, Rob J Leer, Willem M de Vos, Roland J Siezen, Michiel Kleerebezem
Affiliations

Functional analysis of three plasmids from Lactobacillus plantarum

Richard van Kranenburg et al. Appl Environ Microbiol. 2005 Mar.

Abstract

Lactobacillus plantarum WCFS1 harbors three plasmids, pWCFS101, pWCFS102, and pWCFS103, with sizes of 1,917, 2,365, and 36,069 bp, respectively. The two smaller plasmids are of unknown function and contain replication genes that are likely to function via the rolling-circle replication mechanism. The host range of the pWCFS101 replicon includes Lactobacillus species and Lactococcus lactis, while that of the pWCFS102 replicon also includes Carnobacterium maltaromaticum and Bacillus subtilis. The larger plasmid is predicted to replicate via the theta-type mechanism. The host range of its replicon seems restricted to L. plantarum. Cloning vectors were constructed based on the replicons of all three plasmids. Plasmid pWCFS103 was demonstrated to be a conjugative plasmid, as it could be transferred to L. plantarum NC8. It confers arsenate and arsenite resistance, which can be used as selective markers.

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Figures

FIG. 1.
FIG. 1.
Physical and genetic maps of plasmids pWCFS101, pWCFS102, and pWCFS103. The unique XbaI sites used to construct pWCFS104 and pWCFS105 are indicated. The replicon region of pWCFS103 used to construct pWCFS106 is boxed grey.
FIG. 2.
FIG. 2.
Alignment of the double-stranded origins of plasmids pWV01/pFX2 (23) and pLC2 (37) and the putative double-stranded origin of pWCFS102. Dashed arrows indicate the inverted repeats. The arrowheads indicate the nick sites in pFX2 (23). Alignment mismatches are boxed in grey.
FIG. 3.
FIG. 3.
Iteron region of pWCFS103. Dashed arrows indicate the direct and inverted repeats. Mismatches from the consensus sequence are boxed grey. The ribosome binding site (boldface) and start codon (underlined) of repB are indicated.
FIG. 4.
FIG. 4.
Arsenate and arsenite resistance in L. plantarum. Filled diamonds, WCFS1; open diamonds, WCFS2 (arsB); open circles, WCFS3 (cadD); open squares, WCFS4 (traA). Each data point is the mean of duplicate cultures. Error bars indicate the deviation from the mean for single cultures. OD600, optical density at 600 nm.
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References

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