Determination of Luciferase Activity in Arabidopsis seedling

Materials and Reagents

1. Transgenic Arabidopsis seedlings expressing Luciferase fused to promoter of interest (i.e. PHT1;1:Luc in pGWB435 binary vector)
   Note: Vector information can be find at: http://shimane-u.org/nakagawa/pgwb-tables/2.htm.
   
2. KH₂PO₄ (Sigma-Aldrich, catalog number: P5655 )
   
3. Sodium arsenate dibasic heptahydrate (Sigma-Aldrich, catalog number: A6756 )
   
4. 30 mM arsenate (see Recipes)
   
5. 0.36 mM D-Luciferin (Sigma-Aldrich, catalog number: L9504 ) (see Recipes)
   
6. Johnson media (see Recipes)
   

Equipment

1. Growth chamber [16-h-light/8-h-dark regime (24 °C/21 °C)]
   
2. White 96-well plates (Greiner Bio-One GmbH, catalog number: 655074 )
   
3. MicroAmp optical adhesive film (Life Technologies, Applied Biosystem^®, catalog number: 4311971 )
   
4. Luminometer (centro LB 960) (Berthold Technologies)
   

Software

1. Mikro Win 2000 software

Procedure