Transient plant gene expression: a simple and reproducible method based on flowing particle gun.
Successful transient expression of beta-glucuronidase (Gus) and luciferase (Luc) in Nicotiana tabacum leaves was obtained after gene delivery by a simple and inexpensive particle gun. Takeuchi's biolistic process system was adapted to accelerate directly in a helium stream DNA-coated microprojectiles into intact plant leaf tissues. After bombardment of p70-omega-59Gus construction (duplication of the CaMV 35S enhancer), variability inherent to the bombardment procedure was observed in Gus activity measurements between replicate samples. We therefore included an internal standard (35S-Luc) in each bombardment, allowing an accurate, sensitive and reproducible comparison of the effective strength of different Gus-reporter constructs based on Gus/Luc ratio measurements.
Godon
C
C
Laboratoire de Biologie Cellulaire, INRA, Versailles, France.
Caboche
M
M
Daniel-Vedele
F
F
D015870:Gene Expression; D018014:Gene Transfer Techniques; D017343:Genes, Plant; D005821:Genetic Techniques; D005966:Glucuronidase / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D010947:Plants, Toxic; D010957:Plasmids; D015203:Reproducibility of Results; D014026:Nicotiana / Q000235:genetics*
D008156:Luciferases; D005966:Glucuronidase