Download emi12947-sup-0001-si

 marker
(b)
DG 898
DG 898
 marker
(a)
Chromosome
291 kb
pMaD1
pMaD2
pMaD3/4
pMaD5
pMaD6
pMaD7/8
pMaD9
References:
Pradella, S., Allgaier, M., Hoch, C., Päuker, O., Stackebrandt, E., and Wagner-Döbler, I. (2004). Genome
pMaD10
Mu-like prophage
Supplemental Figure S1:
Identification of an excised linear Mu-like phage genome in Marinovum algicola DG898.
(a) Pulsed field gel electrophoresis (PFGE) profile. The PFGE profile based on uncut high
molecular weight genomic DNA. The positions of the DG898 chromosome and ten
plasmids/chromids (pMaD1-10) are indicated. The 11th plasmid of strain DG898 - pMaD11 (5.8kb) - was too small to be detected by PFGE. The dominant band with an estimated size of 34-kb
was supposed to be a linear phage genome in our previous study (Pradella et al., 2010).
Genome sequencing of M. algicola DG898 revealed the presence of a probably intact Mu-like
prophage of 35.5 kb integrated into the chromosome [MALG_02481-MALG_02437; (-)
2,549,072-2,513,591], which is closely related to the Rhodovulum phage RS1 (NC_020866). (b)
Southern blot hybridization. The PFGE gel (Fig. S1a) was hybridized using a DIG-labeled PCRprobe targeting gene MALG_02456 [DG898 chromosome: (-) 2,534,006-2,533,421], which was
annotated as Mu-like FluMu protein gp28. The strong hybridization signal at approximately 34kb confirmed that this band represents the excised genome of the linear Mu-like phage. The
faint hybridization signal with the chromosome furthermore showed that the prophage is still
integrated into the DG898 genome. Hybridization of a second probe deduced from the Mu-like
prophage gene MALG_02444 [(-) 2,525,705-2,525,299; hypothetical protein] gave analogous
signals (data not shown).
Experimental procedures. The PFGE was performed on an 1.2% (w/v) agarose gel with pulse
times of 1–80 s for 45 h at 200 V (6 V/cm). The  low range molecular weight PFGE marker was
used as size standard (New England Biolabs, Beverly, MA, USA). Southern blotting and
hybridization under stringent conditions was done as described by Pradella et al. 2004.
organization and localization of the pufLM genes of the photosynthesis reaction center in
23 kb
phylogenetically diverse marine Alphaproteobacteria. Appl Environ Microbiol 70: 3360-3369.
Pradella, S., Päuker, O., and Petersen, J. (2010). Genome organisation of the marine Roseobacter clade
member Marinovum algicola. Arch Microbiol 192: 115-126.