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pGIAK1, a heavy metal resistant plasmid from an obligate alkaliphilic and halotolerant bacterium isolated from the Antarctic Concordia station confined environment.

Guo S, Mahillon J - PLoS ONE (2013)

Bottom Line: Analysis of the pGIAK1 38,362-bp sequence revealed that, in addition to its replication region, this plasmid contains the genetic determinants for cadmium and arsenic resistances, putative methyltransferase, tyrosine recombinase, spore coat protein and potassium transport protein, as well as several hypothetical proteins.The pGIAK1 replicon region was also shown to be functional in Bacillus thuringiensis, Bacillus subtilis and Staphylococcus aureus, but was only stably maintained in B. subtilis.Finally, using an Escherichia coli - B. thuringiensis shuttle BAC vector, pGIAK1 was shown to display conjugative properties since it was able to transfer the BAC plasmid among B. thuringiensis strains.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Food and Environmental Microbiology, Université Catholique de Louvain, Croix du Sud, Louvain-la-Neuve, Belgium.

ABSTRACT
pGIAK1 is a 38-kb plasmid originating from the obligate alkaliphilic and halotolerant Bacillaceae strain JMAK1. The strain was originally isolated from the confined environments of the Antarctic Concordia station. Analysis of the pGIAK1 38,362-bp sequence revealed that, in addition to its replication region, this plasmid contains the genetic determinants for cadmium and arsenic resistances, putative methyltransferase, tyrosine recombinase, spore coat protein and potassium transport protein, as well as several hypothetical proteins. Cloning the pGIAK1 cad operon in Bacillus cereus H3081.97 and its ars operon in Bacillus subtilis 1A280 conferred to these hosts cadmium and arsenic resistances, respectively, therefore confirming their bona fide activities. The pGIAK1 replicon region was also shown to be functional in Bacillus thuringiensis, Bacillus subtilis and Staphylococcus aureus, but was only stably maintained in B. subtilis. Finally, using an Escherichia coli - B. thuringiensis shuttle BAC vector, pGIAK1 was shown to display conjugative properties since it was able to transfer the BAC plasmid among B. thuringiensis strains.

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Comparison of pGIAK1 and the heavy metal resistant plasmid pWCFS103 from L. plantarum.The displayed bar represents a length of 5,000 basepairs. The conserved regions are blue-shaded, the color intensity indicating the identity levels (from 35 to 100%). Genes that displayed homologies are indicated by the colored boxes shown below. The alignment was made by the Easyfig 2.1 program [49].
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pone-0072461-g003: Comparison of pGIAK1 and the heavy metal resistant plasmid pWCFS103 from L. plantarum.The displayed bar represents a length of 5,000 basepairs. The conserved regions are blue-shaded, the color intensity indicating the identity levels (from 35 to 100%). Genes that displayed homologies are indicated by the colored boxes shown below. The alignment was made by the Easyfig 2.1 program [49].

Mentions: The complete sequence of pGIAK1 was determined to be 38,362 bp, with a G+C content of 37.5%. By BLASTN [48] analysis, 95% of the plasmid DNA sequence had no homologue in the GenBank database. However, it potentially contained 49 ORFs, from which 40 had BLASTP significant similarities to sequences in the GenBank protein database (see Table S1). Based on these homologies, a potential replication region, putative arsenic and cadmium resistance operons, and potential conjugation-related genes could be identified (Figure 2, see below). Using the Easyfig 2.1 software [49], functional similarities could be identified between pGIAK1 and the heavy metal resistant plasmid pWCFS103 [50] from the distantly related Lactobacillus plantarum. As shown in Figure 3, both plasmids contain genes related to conjugation, cadmium and arsenic resistance, although the genes are arranged differently. Their conjugation regions shared no similarities while their ArsB, CadD and RepB proteins shared 68%, 36% and 38% identity, respectively.


pGIAK1, a heavy metal resistant plasmid from an obligate alkaliphilic and halotolerant bacterium isolated from the Antarctic Concordia station confined environment.

Guo S, Mahillon J - PLoS ONE (2013)

Comparison of pGIAK1 and the heavy metal resistant plasmid pWCFS103 from L. plantarum.The displayed bar represents a length of 5,000 basepairs. The conserved regions are blue-shaded, the color intensity indicating the identity levels (from 35 to 100%). Genes that displayed homologies are indicated by the colored boxes shown below. The alignment was made by the Easyfig 2.1 program [49].
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3756968&req=5

pone-0072461-g003: Comparison of pGIAK1 and the heavy metal resistant plasmid pWCFS103 from L. plantarum.The displayed bar represents a length of 5,000 basepairs. The conserved regions are blue-shaded, the color intensity indicating the identity levels (from 35 to 100%). Genes that displayed homologies are indicated by the colored boxes shown below. The alignment was made by the Easyfig 2.1 program [49].
Mentions: The complete sequence of pGIAK1 was determined to be 38,362 bp, with a G+C content of 37.5%. By BLASTN [48] analysis, 95% of the plasmid DNA sequence had no homologue in the GenBank database. However, it potentially contained 49 ORFs, from which 40 had BLASTP significant similarities to sequences in the GenBank protein database (see Table S1). Based on these homologies, a potential replication region, putative arsenic and cadmium resistance operons, and potential conjugation-related genes could be identified (Figure 2, see below). Using the Easyfig 2.1 software [49], functional similarities could be identified between pGIAK1 and the heavy metal resistant plasmid pWCFS103 [50] from the distantly related Lactobacillus plantarum. As shown in Figure 3, both plasmids contain genes related to conjugation, cadmium and arsenic resistance, although the genes are arranged differently. Their conjugation regions shared no similarities while their ArsB, CadD and RepB proteins shared 68%, 36% and 38% identity, respectively.

Bottom Line: Analysis of the pGIAK1 38,362-bp sequence revealed that, in addition to its replication region, this plasmid contains the genetic determinants for cadmium and arsenic resistances, putative methyltransferase, tyrosine recombinase, spore coat protein and potassium transport protein, as well as several hypothetical proteins.The pGIAK1 replicon region was also shown to be functional in Bacillus thuringiensis, Bacillus subtilis and Staphylococcus aureus, but was only stably maintained in B. subtilis.Finally, using an Escherichia coli - B. thuringiensis shuttle BAC vector, pGIAK1 was shown to display conjugative properties since it was able to transfer the BAC plasmid among B. thuringiensis strains.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Food and Environmental Microbiology, Université Catholique de Louvain, Croix du Sud, Louvain-la-Neuve, Belgium.

ABSTRACT
pGIAK1 is a 38-kb plasmid originating from the obligate alkaliphilic and halotolerant Bacillaceae strain JMAK1. The strain was originally isolated from the confined environments of the Antarctic Concordia station. Analysis of the pGIAK1 38,362-bp sequence revealed that, in addition to its replication region, this plasmid contains the genetic determinants for cadmium and arsenic resistances, putative methyltransferase, tyrosine recombinase, spore coat protein and potassium transport protein, as well as several hypothetical proteins. Cloning the pGIAK1 cad operon in Bacillus cereus H3081.97 and its ars operon in Bacillus subtilis 1A280 conferred to these hosts cadmium and arsenic resistances, respectively, therefore confirming their bona fide activities. The pGIAK1 replicon region was also shown to be functional in Bacillus thuringiensis, Bacillus subtilis and Staphylococcus aureus, but was only stably maintained in B. subtilis. Finally, using an Escherichia coli - B. thuringiensis shuttle BAC vector, pGIAK1 was shown to display conjugative properties since it was able to transfer the BAC plasmid among B. thuringiensis strains.

Show MeSH
Related in: MedlinePlus