HighlightsHeterologous sfCherry protein was expressed in N. salina for the first time.N. salina was transformed by particle bombardment.Integration site of the transgene on the genome was determined by RESDA PCR.Expression of sfCherry was confirmed by a western blotting and confocal microscopy.

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The influence of nitrogen (N) deficiency on the cell growth and intracellular lipid production of the alga Botryococcus braunii UTEX 572 was investigated. Biomass concentration and lipid content of B. braunii cultivated in modified Chu-13 medium containing 0.04, 0.37, and 3.66 mM nitrate were 0.23-0.38 g L −1 and 36-63% of dry cell weight, respectively. The specific growth rate of B. braunii reached a constant of 0.185 day −1 during cultivation with an initial nitrate feed of 3.66 mM. The maximum lipid content of B. braunii was 63% with 0.04 mM nitrate. However, the maximum lipid productivity of 0.019 g L −1 day −1 was achieved with 0.37 mM nitrate. The level of oleic acid, an important component of biodiesel, was higher at 86% of the total fatty acids under N-limited conditions (0.04 mM nitrate) compared to 69% under N-sufficient conditions (3.66 mM nitrate). Furthermore, expression of the stearoyl-ACP desaturase gene (sad) encoding a stearoyl-ACP desaturase involved in the synthesis of oleic acid was 2.6-fold higher under N-limited conditions than under N-sufficient conditions.

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A simple and rapid method is presented for the preparation of RNA from various cyanobacteria. Unlike other methods that require a lysis solution, lysozymes, or proteinase K, the proposed method, called the bead-phenol-chloroform (BPC) method, uses silica/zirconia beads, phenol, and chloroform to break the cells and extract RNA more efficiently. Experiments confirm that the BPC method can successfully isolate total RNA from various cyanobacterial strains without DNA contamination, and the extracted RNA samples have a relatively high purity, concentration, and yield. Furthermore, the BPC method is more rapid, simple, and economical when compared with previously reported methods.