Optimization of Electro-Transformation Conditions for the Transformation Human Lactoferrin of Desert Chlorella

Abstract:

To establish an efficient genetic transformation protocol for Desert Chlorella. It was optimized methods for electroporation conditions and related parameters. Human lactoferrin gene regard as reference gene of optimization of electro-transformation conditions. The various parameters of cell viability and transfection efficiency were determined with single factor experiments.Then，the optimal electroporation conditions were obtained through orthogonal test. Results shows the optimal composition of electroporation conditions：breakdown voltage 1 400 V，cell cycle 8day，osmosis buffer 0.2 mol/L，osmosis time 0 h，plasmid 6 g/mL，electroporation buffer 0.4 mol/L. Under the optimal composition of electroporation conditions，the detected results of cell viability is 51.30%，and fluorescence positive rate is 52.54%，and PCR positive rate is 42.30%.