Journal: PLOS ONE

Article Title: A novel SfaNI-like restriction-modification system in Caldicellulosiruptor extents the genetic engineering toolbox for this genus

doi: 10.1371/journal.pone.0279562

Figure Lengend Snippet: List of strains used and constructed in this study.

Article Snippet: Caldicellulosiruptor bescii DSM 6725 , DSMZ.

Techniques: Construct

Journal: PLOS ONE

Article Title: A novel SfaNI-like restriction-modification system in Caldicellulosiruptor extents the genetic engineering toolbox for this genus

doi: 10.1371/journal.pone.0279562

Figure Lengend Snippet: Digestion of genomic DNA of type strains of Caldicellulosiruptor species with the restriction endonucleases HaeIII and SfaNI.

Article Snippet: Caldicellulosiruptor bescii DSM 6725 , DSMZ.

Techniques:

Journal: PLoS ONE

Article Title: Methylation by a Unique α-class N4-Cytosine Methyltransferase Is Required for DNA Transformation of Caldicellulosiruptor bescii DSM6725

doi: 10.1371/journal.pone.0043844

Figure Lengend Snippet: Strains/plasmids used and constructed in this study.

Article Snippet: DSM 6725 , Caldicellulosiruptor bescii wild type , DSMZ .

Techniques: Construct, Marker, Expressing, Plasmid Preparation

Journal: PLoS ONE

Article Title: Methylation by a Unique α-class N4-Cytosine Methyltransferase Is Required for DNA Transformation of Caldicellulosiruptor bescii DSM6725

doi: 10.1371/journal.pone.0043844

Figure Lengend Snippet: (A) A diagram of the pyrBCF locus in the C. bescii chromosome. The line above the diagram depicts the extent of the spontaneous deletion in the Δ pyrBCF strain. pDCW70 contains the wild type pyrBCF alleles with an engineered KpnI site used to select marker replacement of the deletion depicted with primers used to confirm the structure of the chromosome in the transformant. B) Δ pyrBCF electrocompetent cells after electro-pulse with no DNA added plated onto defined medium + uracil (top plate), Δ pyrBCF electrocompetent cells after electro pulse with unmethylated pDCW70 DNA plated onto defined medium w/o uracil (middle plate), Δ pyrBCF electrocompetent cells after electro-pulse with M.CbeI methylated pDCW70 DNA plated onto defined medium w/o uracil (bottom plate). (C) PCR products amplified using primers DC163 and DC188 (upper gel) digested with KpnI (lower gel). M: 1kb DNA Ladder (NEB). lane 1: amplified from wild type cells (3.2 kb); lane 2: amplified from Δ pyrBCF (1.63 kb); lane 3: amplified from transformants (1.9 and 1.3 kb cleavage products).

Article Snippet: DSM 6725 , Caldicellulosiruptor bescii wild type , DSMZ .

Techniques: Marker, Methylation, Amplification

Journal: PLoS ONE

Article Title: Methylation by a Unique α-class N4-Cytosine Methyltransferase Is Required for DNA Transformation of Caldicellulosiruptor bescii DSM6725

doi: 10.1371/journal.pone.0043844

Figure Lengend Snippet: Strains/plasmids used and constructed in this study.

Article Snippet: DSM 6725 , Caldicellulosiruptor bescii wild type , DSMZ .

Techniques: Construct, Marker, Expressing, Plasmid Preparation

Journal: PLoS ONE

Article Title: Methylation by a Unique α-class N4-Cytosine Methyltransferase Is Required for DNA Transformation of Caldicellulosiruptor bescii DSM6725

doi: 10.1371/journal.pone.0043844

Figure Lengend Snippet: Predicted functional domains of M.CbeI and sequence alignments of conserved motifs of three M.CbeI homologues from members of Caldicellulosiruptor species as well as DmtB from Anabaena variabilis , which also contains an M.CbeI homologue showing unique features of this methyltransferase.

Article Snippet: DSM 6725 , Caldicellulosiruptor bescii wild type , DSMZ .

Techniques: Functional Assay, Sequencing

Journal: PLoS ONE

Article Title: Methylation by a Unique α-class N4-Cytosine Methyltransferase Is Required for DNA Transformation of Caldicellulosiruptor bescii DSM6725

doi: 10.1371/journal.pone.0043844

Figure Lengend Snippet: Strains/plasmids used and constructed in this study.

Article Snippet: C. bescii DSM 6725 was inoculated into 10 ml of modified DSMZ 516 medium and grown anaerobically at 60°C for 24 h. Cells were harvested at 18,000 ×g for 5 min, washed twice with 1 x base salts , resuspended in 1 ml of 1 x base salts and plated by mixing 100 µl of cell suspension with 4 ml of 0.3% agar and overlaying onto defined modified DSMZ 516 medium (no yeast extract or casein) supplemented with 20 µM uracil and 8 mM 5-FOA (US Biologicals, Swampscott, MA).

Techniques: Construct, Marker, Expressing, Plasmid Preparation