Fig. 1

Transformation of Nannochloropsis oceanica using Tn5 transposome. a The CMV promoter allows expression of the Zeocin resistance gene in mammalian cells. The EM7 promoter is a synthetic promoter based on the bacteriophage T7 promoter for expression of the Zeocin resistance factor in E. coli. Shble is the gene encoding antibiotic resistance to Zeocin. Mosaic ends (ME) are recognition sites for the binding of the transposase Tn5. FF Zeo and Rev Zeo primers amplify the coding region of the Shble resistance gene. Primers FF pMOD and Rev pMOD amplify the complete construct from the pMOD vector. b PCR of colonies of transformed (T) and non-transformed (−/T) N. oceanica clones using specific primers for Zeocin resistance (374 pbs), control (+) plasmid pMOD-Zeo. C) Growth of mutant clones of transposon transformed-Nannochloropsis oceanica on agar plates supplemented with zeocin (2 µg/ml)