Fig. 2

Insertional mutagenesis and FACS-based screening to isolate high lipid-accumulating N. salina. a Schematic map of pNsShble. b FACS-based analysis for screening of insertional mutagenesis library. Microalgal cells were stained with BODIPY to detect the intracellular lipid in vivo. The stained samples were analyzed by using FACS and each of the mutants was arranged in the order of relative fluorescence intensity emitted from BODIPY. The X-axis represents the total analyzed single mutants, and the Y-axis represents the fluorescence detected from each mutants. The relative fluorescence intensity was calculated using mean value of four independent experiments. WT (black) and Mut68 (green) were indicated. c Confirmation of gene integration of Mut68 by performing PCR. M, marker; lane 1, WT; lane 2, Mut68. d Analysis of inserted copy number of pNsShble by performing Southern blot analysis. Lane 1, WT; lane 2, Mut68