Gene stacking is a method used in biotechnology by which multiple genes can be placed at a single genomic site, thereby simplifying plant breeding. In this approach, DNA nucleases are used for excising selectable marker genes (SMG), which are the unneeded components of transgenic plants. The goal of this project is to evaluate the effectiveness of the nuclease I-SceI in excising DNA in plants. Specifically, this study tests heat-inducible I-SceI through the use of a heat-shock promoter (HS) in order to control SMG excision by heat application. The DNA plasmid containing a visual marker gene flanked by I-SceI target sites and the heat-inducible I-SceI gene has been created and confirmed. Arabidopsis thaliana plants have been transformed with the plasmid, which will be used for testing the efficiency of HS:I-SceI in excising DNA from plant genomes.
Pruett, Elliot E.; Nandy, Soumen; and Srivastava, Vibha
"Efficiency of the nuclease I-SceI in excising selectable marker genes from the plant genome,"
Discovery, The Student Journal of Dale Bumpers College of Agricultural, Food and Life Sciences. University of Arkansas System Division of Agriculture. 17:85-90.
Available at: https://scholarworks.uark.edu/discoverymag/vol17/iss1/14