Optimization of Parameters for GUS Gene Transformation of Porphyra yezoensis by Particle Bombardment

摘要

We optimized the biological and physical parameters for DNA delivery into thalli of the red alga Porphyra yezoensis using a particle bombardment device. The efficiency of transformation was determined using the p-glucuronidase (GUS) assay. The optimalhelium pressure, distance of tungsten particle flight, and ratio of DNA to tungsten particles were 23 kgf/cm~2, 8 cm, and 5 mu g/mg tungsten, respectively. During bombardment, osmotic treatment with a mixture of 0.6 M mannitol and sorbitol increased theefficiency of GUS transformation. After 2 days, the blue color indicating GUS activity was observed using a histochemical assay.