MOESM1 of Wavelength shift strategy to enhance lipid productivity of Nannochloropsis gaditana

Additional file 1: Figure S1. Configuration of PBR with dye-filled layer

Protein C is the 31 kDa fragment of FibA.

<p><b>A and B</b>) Immunoblot analysis using anti-Protein C polyclonal antibodies (pAb) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028968#pone.0028968-McCleary1" target="_blank">[9]</a> (left) and anti-FibA 2105 monoclonal antibodies (mAb) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028968#pone.0028968-Kearns1" target="_blank">[13]</a> (right). Black arrows: ∼66 kDa band previously assigned to FibA <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028968#pone.0028968-Kearns1" target="_blank">[13]</a>. Grey arrows: ∼31 kDa band previously assigned to FibA <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028968#pone.0028968-Kearns1" target="_blank">[13]</a> and to Protein C <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028968#pone.0028968-McCleary1" target="_blank">[9]</a>. <b>A</b>) Total cell lysates prepared from equal numbers of cells from vegetative cultures (0 hours development) of <i>dsp</i> (DK3470) and wild type (DZ2) cells, and <i>dsp</i> cultures developing for 24 hours on nutrient-limited CF agar plates. <b>B</b>) FibA is present only in the aggregated cell fraction. Wild type (DZ2) and <i>fibA</i> (PH1018) cells were developed under submerged culture for 24 hours. Cell lysates were prepared from aggregated cell fractions (P) and supernatant cell fractions (S) as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028968#pone-0028968-g003" target="_blank">Fig. 1</a>.</p

The <i>fibA</i> promoter region is active in both cell fractions.

<p><b>A</b>) Coomassie stain (left), and anti-Protein C (<i>aka</i> FibA) (middle), or anti-mCherry (right) immunoblot analysis of aggregated (P) and supernatant (S) cell fractions harvested from strain PH1019 (DZ2 <i>attB</i>::P_fibA-mCherry) developed for 24 hours under submerged culture. Supernatant and aggregated cell lysates were prepared from equal numbers of cells. Black and grey arrows indicate the ∼66 kDa and ∼31 kDa bands previously attributed to FibA and FibA/Protein C, respectively. <b>B</b>. Distribution of individual cell mCherry fluorescence intensities recorded from the samples above. Background subtracted intensity measurements of ≥250 cells from each fraction were recorded. The distribution of intensity measurements (bin size 50 relative intensity values) is displayed as a histogram for the aggregated (pelleted fraction) and supernatant cell fractions as indicated. Histograms were generated using Origin (ver. 6.1) data analysis and graphing software (Northampton, MA, USA). <b>A and B</b>. Results from one assay are shown, but triplicate biological repetitions produced identical results.</p

Sporulation efficiencies of wild type and <i>fibA</i> cells developed in submerged culture or on CF agar.

a<p>percent of wild type spores produced at 120 hours of development.</p>b<p>percent of wild type germinating spores.</p>c<p>none detected; <1×10⁵ spores per spot (<0.21% of wild type).</p

Protein C accumulation is heterogeneous.

<p><b>A</b>. Anti-Protein C <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028968#pone.0028968-McCleary1" target="_blank">[9]</a> immunoblot analysis of wild type (<i>M. xanthus</i> strain DZ2) cells grown on the surface of a Petri plate in vegetative (CYE) media. Cells were harvested, and cells in aggregates were pelleted at 50 x g for 5 min. Each lane contains lysate from 4.3×10⁷ cells harvested from the supernatant (S) or aggregated cell pellet (P) fractions. FB: Control demonstrating the Protein C accumulation in 4.3×10⁷ cells isolated from fruiting bodies formed after 48 hours of development. B. Anti-PilA <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028968#pone.0028968-Wu1" target="_blank">[17]</a> (top panel) and anti-PilC <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028968#pone.0028968-Bulyha1" target="_blank">[16]</a> (bottom panel) immunoblot of the supernatant (S) or pellet (P) fractions from A. Protein C was detected exclusively in the pellet cell fraction whereas PilA and PilC were equally represented in both cell fractions.</p

Mass spectrometry maps Protein C to FibA.

<p>Domain architecture of the 744 amino acid (aa) FibA preprometalloprotease as predicted by SMART (<a href="http://smart.embl-heidelberg.de" target="_blank">http://smart.embl-heidelberg.de</a>) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028968#pone.0028968-Schultz1" target="_blank">[54]</a>. ss: signal sequence (aa 1–24); FTP: Fungalysin/Thermolysin Propeptide PFAM domain (aa 100–149); Peptidase _M4/M4_C: peptidase family M4 and M4-Cterminal PFAM domains (aa 218–518); PPC: Bacterial pre-peptidase C-terminal PFAM domain (aa 544–614 and 638–724). Black lines correspond to the length and sequence position of the peptides identified from mass spectrometry analysis of Protein C isolated from the spore coat (above schematic) or by immune-precipitation of the aggregated cell fraction using anti-Protein C sera (below schematic).</p

MOESM1 of Isolation, phenotypic characterization and genome wide analysis of a Chlamydomonas reinhardtii strain naturally modified under laboratory conditions: towards enhanced microalgal biomass and lipid production for biofuels

Additional file 1: Table S1. Specific growth rate of CC-124 in year 2010, 2013, and 2015

MOESM2 of Isolation, phenotypic characterization and genome wide analysis of a Chlamydomonas reinhardtii strain naturally modified under laboratory conditions: towards enhanced microalgal biomass and lipid production for biofuels

Additional file 2: Figure S1. Cell size distribution of CC-124L and CC-124H in liquid TAP medium

Notice des meilleurs livres nouveaux, traitant des établissemens ou réformes utiles à l'humanité, au bien d'un Etat, et au bonheur des hommes en général, et particulièrement sur les hôpitaux ; chez Royez, libraire, quai des Augustins, à Paris

[Catalogue de libraire. Paris. Royez, Jean-François. 1788]Avec mode text

Boletín de Segovia: Número 78 - 1923 junio 29

Copia digital. Madrid : Ministerio de Cultura. Subdirección General de Coordinación Bibliotecaria, 200