Assay of Firefly Luciferase in Transformed Maize Tissues

Abstract

This protocol describes the assay of firefly luciferase enzyme activity in homogenates of maize cells transiently transformed with plasmid constructs able to express the firefly luciferase gene. A variety of intact plant tissues or cultured maize cell lines may be used for the detection of firefly luciferase activity following gene transfer. Transformation techniques are described in other chapters. The general approach is to homogenize the tissue, centrifuge down the debris, and determine the amount of luciferase by measuring the light output when the extract is mixed with the substrates luciferin and ATP in a luminometer. This assay is very sensitive and has been used to detect luciferase expression in a very low percentage of the total cell population.