Microalgal Colony Blot: a simple and rapid method for direct detection of recombinant protein production in microalgae colonies

Supporting Data for Microalgal colony blot: a simple and rapid method for direct detection of recombinant proteins in microalgae Torres-Tiji Y., Fields F., Mayfield S. P. (2025)PLOS ONE Contents Plasmid Sequences GenBank (.gbk) and SnapGene (.dna) formats are provided for all plasmids. mCloverGFP expression plasmid (.gbk, .dna)Ble-mCloverGFP fusion expression vector used for primary screening ICAM1 expression plasmid (.gbk, .dna)Ble-ICAM1 fusion expression vector (FLAG-tagged) CtxB-RBD expression plasmid (.gbk, .dna)Cholera toxin subunit B fused to SARS-CoV-2 spike protein RBD RBD-mClover secreted expression plasmid (.gbk, .dna)RBD-mClover fusion with secretion signal RBD-mClover ER-retained expression plasmid (.gbk, .dna)RBD-mClover fusion with ER retention signal Flow Cytometry Data Flow cytometry standard (.fcs, version 3.0) files acquired on a CytoFLEX (Beckman Coulter) using CytExpert 2.4. Wild-type controlsWT1.fcs, WT2.fcs High-expressing clonesH1.fcs, H2.fcs, H3.fcs Medium-expressing clonesM1.fcs, M2.fcs, M3.fcs Low-expressing clonesL1.fcs, L2.fcs, L3.fcs Gated cell populations (.xlsx)Exported scale values for gated populations used in analysis Acquisition settings (identical for all samples): FL1 (GFP FITC): Gain 60; Ex 488 nm; Em 525/40 nm FL3 (Auto APC): Gain 600; Ex 638 nm; Em 660/10 nm FSC/SSC: Gain 500; Threshold 50,000 / 10,000 Original Blot Images Uncropped, unadjusted original images provided in compliance with PLOS ONE data-availability requirements. Figure 1 Panel A – original picture (.JPG) Panel B – original picture (.JPG) Panel C – original picture (.JPG) Panel D – original picture (.JPG)Microalgal colony blot development time series Figure 2 Original picture (.JPG)western blot validation of selected clones Figure 5 Panel A – ICAM1 MCB algae original picture (.JPG)ICAM1-expressing colonies on plate Panel B – ICAM1 MCB original picture (.JPG)ICAM1 microalgal colony blot Panel C – RBD MCB algae original picture (.JPG)RBD-expressing colonies on plate Panel D – RBD MCB original picture (.JPG)RBD microalgal colony blot Panel E – MCB C. pacifica algae original picture (.JPG)C. pacifica colonies on plate Panel F – MCB C. pacifica original picture (.JPG)C. pacifica microalgal colony blot Figure S2 Anti-RBD western blot original picture (.JPG) Anti-CtxB western blot original picture (.JPG) File Formats .gbk — GenBank format (SnapGene, Benchling, ApE, UGENE) .dna — SnapGene format (annotations preserved; free viewer available) .fcs — Flow Cytometry Standard 3.0 (FlowJo, CytExpert, FCS Express) .xlsx — Microsoft Excel format .JPG — JPEG image format Contact Corresponding author:Yasin Torres-Tiji Colorado School of Mines / UC San Diego [e-mail]/[e-mail]