Identification of the genetic rearrangement in the suppressor strain.

<p>PCR amplifications were carried out to narrow down the site of possible genetic rearrangement in the <i>rpoE</i>-<i>nadB</i> region of the chromosome. (<b>A</b>, <b>B</b>) Approximate drawings showing the <i>rpoE</i> and <i>nadB</i> genes from the wild-type (<b>A</b>) and suppressor (<b>B</b>) strains, as well as the positions of primers (numbered 1 to 4), their orientations, and size of the amplified DNA fragments. (<b>C</b>) Agarose gels showing the results of PCR amplifications from the wild-type and suppressor strains. Numbers 1 to 4 refer to the primers shown in (<b>A</b>) and (<b>B</b>).</p

The suppressor mutation reverses the vesiculation phenotype of a Δ<i>degP</i> strain.

<p>Cultures were grown at 39°C in Luria broth for five hours, after which membranes and vesicles were prepared as described in the Experimental Procedure section. Samples were analyzed by SDS-PAGE and proteins were visualized after Coomassie blue staining. Abbreviations: mem, membrane; ves, vesicle; <i>sup</i>⁺, the unknown suppressor mutation.</p

Effects of Δ<i>degP</i> and <i>rpoE3</i> mutations on RybB::LacZ (A) and CpxP::LacZ (B) activities.

<p>For each strain, two independent cultures were grown at 39°C to a mid-log phase and used for β-galactosidase assays. LacZ activities are relative to the wild-type strain. The relevant genotypes of the strains are labeled at the bottom.</p

Growth curves of bacterial cultures grown at 39°C.

<p>Wild type and Δ<i>degP</i> cells, with or without the suppressor mutation, were grown overnight at 30°C. Next day, overnight cultures were diluted to 1∶100 in flasks containing fresh, pre-warmed, Luria broth and growth was resumed at 30°C and 39°C. OD₆₀₀ was measured from bacterial samples withdrawn every 30 minutes. Only 39°C growth curves, obtained from two independent experiments, are shown. All strains grew almost identically at 30°C.</p

MALDI-TOF spectra and pseudo-gels of simple mixtures containing two or three individual microalgae.

<p>Representative spectra of samples of <i>Chlorella vulgaris</i> UTEX 395 (a), <i>Scenedesmus acutus</i> LRB-AP 401 (b), <i>Chlorella sorokiniana</i> UTEX 1230 (c), a mixture of <i>C</i>. <i>vulgaris</i> UTEX 395 and <i>S</i>. <i>acutus</i> LRB-AP 401 (d), and a mixture of all three microalgae (e).</p

Peaks observed in spectra of individual microalgae and simple mixtures.

<p>M = peak from individual microalga that was observed in one or both <u>m</u>ixtures</p><p>P = <u>p</u>eak from individual microalga that was not observed in either mixture</p><p>S = mixture-<u>s</u>pecific peak</p><p>Peaks observed in spectra of individual microalgae and simple mixtures.</p

Representative MALDI-TOF spectra and pseudo-gels of microalgae strains of the same species.

<p>Three <i>C</i>. <i>vulgaris</i> spectra (a–c) exhibit strain-level differences: UTEX 395 (a), UTEX 259 (b), and LRB-AZ 1201 (c). Three <i>N</i>. <i>oceanica</i> spectra (d–f) also exhibit strain-level differences: CCMP 531 (d), IMET-1 (e), and CCAP 849/10 (f).</p

Microalgae species and strains used in this study.

<p>¹ Carolina 152069 was identified by Carolina Biological as <i>Chlorella spp</i>.</p><p>² Carolina 152280 was identified by Carolina Biological as <i>Haematococcus spp</i>.</p><p>Microalgae species and strains used in this study.</p

Similarity-based dendrograms representing spectra (a) and 18S rDNA sequences (b) of 31 microalgae.

<p>Class names are listed on the right to facilitate comparison between the two dendrograms. The 18S rDNA sequence-based dendrogram (b) shows grouping of the samples up to the class level. Spectra (a) were clustered using the UPGMA algorithm, while sequences (b) were clustered using the neighbor joining algorithm.</p

MOESM2 of A type-I diacylglycerol acyltransferase modulates triacylglycerol biosynthesis and fatty acid composition in the oleaginous microalga, Nannochloropsis oceanica

Additional file 2: Table S1. DGAT protein sequences used for the construction of phylogenetic tree in additional file 1: Figure S3. Table S2. Primers used in the present study. Underlined sequences designate the restriction enzyme sites. The sequences in box indicate the linker fragment introduced before GFP coding sequence