TRANSIENT PLANT GENE-EXPRESSION - A SIMPLE AND REPRODUCIBLE METHOD BASED ON FLOWING PARTICLE GUN

Successful transient expression of beta-glucuronidase (Gus) and lucife rase (Luc) in Nicotiana tabacum leaves was obtained after gene deliver y by a simple and inexpensive particle gun. Takeuchi's biolistic proce ss system was adapted to accelerate directly in a helium stream DNA-co ated microprojectiles into intact plant leaf tissues. After bombardmen t of p70-OMEGA-59Gus construction (duplication of the CaMV 35S enhance r), variability inherent to the bombardment procedure was observed in Gus activity measurements between replicate samples. We therefore incl uded an internal standard (35S-Luc) in each bombardment, allowing an a ccurate, sensitive and reproducible comparison of the effective streng th of different Gus-reporter constructs based on Gus/Luc ratio measure ments.