r centenaria  (ATCC)

Journal: BMC Microbiology

Article Title: The photosensor protein Ppr of Rhodocista centenaria is linked to the chemotaxis signalling pathway

doi: 10.1186/1471-2180-10-281

Figure Lengend Snippet: Domain structure of the Ppr photosensor protein of R. centenaria . The Ppr protein consists of a photoactive yellow protein domain (Pyp; residues 1-135) which carries the blue light absorbing chromophore p -hydroxycinnamic acid, a central bacteriophytochrome bilin binding domain (residues 136-601) with the red light absorbing biliverdin chromophore, and a histidine kinase domain (Pph; residues 602-884). The truncated Pph protein consists of the histidine kinase domain (residues 602-884). In the mutant Pph H670A the putative autophosphorylated histidine residue (H670) is replaced by an alanine.

Article Snippet: R. centenaria (ATCC 43720) was obtained from the culture collection. (For anaerobic photosynthetic growth R. centenaria was cultured in screw cap bottles filled to the top with PYVS medium [ ] and illuminated by an 80 W tungsten bulb (Concentra, Osram, Germany) at 42°C.

Techniques: Binding Assay, Mutagenesis

Journal: BMC Microbiology

Article Title: The photosensor protein Ppr of Rhodocista centenaria is linked to the chemotaxis signalling pathway

doi: 10.1186/1471-2180-10-281

Figure Lengend Snippet: Interaction between Pph and the chemotactic protein Rc-CheW. (A) The binding of the histidine kinase domain Pph and CheW was analyzed in pull-down assays. R. centenaria 6his-Rc-CheW was expressed in E. coli C41 cells and purified. The Pph protein was translated in vitro in the presence of [ 35 S]-methionine (lane 1 and 4). Rc-CheW was added (50 μg) to the reaction and incubated at 37°C. The sample was applied to a Cu-Sepharose column and after washing the bound complexes were eluted (lanes 3 and 6). The fractions were analysed by phosphorimaging. The in vitro translating protein extracts are shown in lanes 1 and 4, the final wash steps in lanes 2 and 5 and the elution fractions in lanes 3 and 6, respectively. The co-elution rate was calculated and is indicated. The positions of molecular weight markers are indicated. (B) The binding of the Pph protein and Rc-CheW was analysed in the presence of ATP. The Pph protein was translated and Rc-CheW was added as described in (A). ATP or apyrase was added to each reaction as indicated and the samples were analysed as described in (A). The co-elution rate was calculated and is indicated in % as bound Pph protein.

Article Snippet: R. centenaria (ATCC 43720) was obtained from the culture collection. (For anaerobic photosynthetic growth R. centenaria was cultured in screw cap bottles filled to the top with PYVS medium [ ] and illuminated by an 80 W tungsten bulb (Concentra, Osram, Germany) at 42°C.

Techniques: Binding Assay, Purification, In Vitro, Incubation, Co-Elution Assay, Molecular Weight

Journal: BMC Microbiology

Article Title: The photosensor protein Ppr of Rhodocista centenaria is linked to the chemotaxis signalling pathway

doi: 10.1186/1471-2180-10-281

Figure Lengend Snippet: Oligomeric state of the histidine kinase Pph. (A) Alignment of the dimerization domains of the Pph protein from R. centenaria and EnvZ from E. coli . The identity was 27% whereas the similarity was calculated with about 57%. The alignment was performed with the Clustal X software. (B) Purified Pph was analysed by gel filtration on a Sephadex G-200 column. Aliquots of the elution fractions (39-48) were separated by SDS-PAGE and blotted on a nitrocellulose membrane. The Pph protein was detected with a conjugate raised against the C-terminal StrepTag II. The position of the Pph protein is indicated. The following proteins werde used as molecular weight markers: β-amylase (200 kDa), alcohol dehydrogenase (150 kDa), albumin (66 kDa), carboanhydrase (29 kDa) and cytochrome c (12 kDa) were used.

Article Snippet: R. centenaria (ATCC 43720) was obtained from the culture collection. (For anaerobic photosynthetic growth R. centenaria was cultured in screw cap bottles filled to the top with PYVS medium [ ] and illuminated by an 80 W tungsten bulb (Concentra, Osram, Germany) at 42°C.

Techniques: Software, Purification, Filtration, SDS Page, Molecular Weight

Journal: BMC Microbiology

Article Title: The photosensor protein Ppr of Rhodocista centenaria is linked to the chemotaxis signalling pathway

doi: 10.1186/1471-2180-10-281

Figure Lengend Snippet: Protein complexes containing Pph isolated from R. centenaria . The Pph protein C-terminally fused to a strep-tag was expressed in R. centenaria and bound to a streptactin-Sepharose column. The elution fractions were analyzed by SDS-PAGE, silver staining (A) and Western blot with antibodies to strep-tag II (B) or to Rc-CheW (C), respectively. The crude protein extract (lanes 1 and 4), the last washing step (lanes 2 and 5) as well as the elution step (lanes 3 and 6) are shown. The positions of molecular weight markers are indicated.

Article Snippet: R. centenaria (ATCC 43720) was obtained from the culture collection. (For anaerobic photosynthetic growth R. centenaria was cultured in screw cap bottles filled to the top with PYVS medium [ ] and illuminated by an 80 W tungsten bulb (Concentra, Osram, Germany) at 42°C.

Techniques: Isolation, Strep-tag, SDS Page, Silver Staining, Western Blot, Molecular Weight

Journal: Viruses

Article Title: In the Search of Marine Pestiviruses: First Case of Phocoena Pestivirus in a Belt Sea Harbour Porpoise

doi: 10.3390/v14010161

Figure Lengend Snippet: Map and histogram illustrating collection sites (green dots) and distribution of 77 harbour porpoises screened per year between 2007 and 2019. The red star indicates the collection site of the Phocoena pestivirus (PhoPeV)-positive harbour porpoise (ID: 43720 and collection date: 4 February 2011).

Article Snippet: The genome sequence of the PhoPeV strain 43720 detected in lung, spleen and ovary tissue samples of a harbour porpoise collected in Denmark (Zealand) in 2011 (sample ID 43720; ; accession number: OK272505) was determined by next-generation sequencing on an Illumina HiSeq as previously described [ , ].

Techniques: