Optimization of luciferase activity in a tomato transient assay system |
| |
Authors: | Thianda Manzara Svetlana Tarchevskaya Jonathon O. Narita |
| |
Affiliation: | (1) Laboratory for Molecular Biology, Department of Biological Sciences (M/C 067), University of Illinois at Chicago, 840 West Taylor Street, 60607-7020 Chicago, Illinois, USA |
| |
Abstract: | The gene encoding firefly luciferase is a commonly used reporter for transient expression assays in plants. We have found that the concentration of buffers normally used in luciferase assays is too low to adequately buffer acidic plant organs. This results in low apparent luciferase activity as well as high variability among replicates. In a transient assay system based on particle bombardment of ripe tomato fruit, luciferase activity driven by the 35S promoter of cauliflower mosaic virus was increased as much as 130 fold by increasing the concentration of the buffer from 50 mM to 300 mM. Using 300 mM buffer, expression levels of luciferase driven by three different plant promoters were found to reflect expression patterns in intact plants. |
| |
Keywords: | luciferase reporter gene tomato transient assay |
本文献已被 SpringerLink 等数据库收录! |