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Using the VersaFluor Fluorometer to Quantitate GUS Expression in Plant Tissues Bombarded With the Biolistic PDS-1000/He System

as an internal control to reduce variability in all our particle bombardment experiments (Figure 1A). The expression results (Figure 2B) are reported as the ratio of luciferase to GUS activity normalized to 100% for BKm3 expression in abscission zones. Expression of luciferase from the test promoter, the BKm3 construct (Figure 1A), relative to GUS expression from D35S (i.e., luciferase to GUS ratio) in stems and petioles is approximately 10% of the expression in abscission zones (Figure 2B). This pattern of expression is very similar to that for native cellulase activity measured in the same extracts collected from particle bombarded stems, petioles and leaf abscission zones (LAZ) (Figure 2A).

The VersaFluor fluorometer is inexpensive and easy to use. It has the necessary sensitivity to reproducibly measure transient GUS activity in particle gun bombarded plant tissues. In addition, using the Hoechst 33258 dye to quantitate DNA by the VersaFluor fluorometer is reliable, simple, and accurate.


References
1. Jefferson, R. A., Kavanagh, T. A. and Bevan, M. W., EMBO 6, 3901-3907 (1987).

2. Kay, R., Chan, A., Daly, M. and McPherson, J., Science, 236, 1299-1302 (1987).

3. Koehler, S. M., Matters, G. L., Nath, P., Kemmerer, E. C. and Tucker, M. L., Plant Molec. Biol., 31, 595-606 (1996).

4. Ow, D. W., Wood, K. V., DeLuca, M., DeWet, J. R., Helinski, D. R., and Howell, S. H., Science, 234, 856-859 (1986).


Triton is a trademark of Rohm and Haas. Biolistic is a trademark of E.I. Du Pont de Nemours and Co. Biolistic technology is exclusively licensed to Bio-Rad Laboratories.


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