Submitted to: Plant Molecular Biology Reporter
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: March 29, 2006
Publication Date: March 31, 2006
Citation: Thilmony, R.L., Guttman, M.E., Chiniquy, D., Blechl, A.E. 2006. Pgpro 1, a novel binary vector for monocot promoter characterization. Plant Molecular Biology Reporter. 24:1-13, March 2006 Interpretive Summary: In order to make genetic engineering of crop plants more precise and more acceptable to the general public, it is important for plant researchers to have access to promoters which express genes only in the tissues and organs where it is needed. To characterize such elements, we have built the vector, “pGPro1”, with several useful properties. Most importantly, pGPro1 allows researchers to study promoter function without inference from other nearby sequences. We show that pGPro1 works in onion and rice cells and that it can be used to study promoters in transformed rice plants. This new vector will be very useful to researchers who want to find promoter sequences with tissue-specific expression patterns, which, in turn can be utilized to confine foreign gene products to non-edible parts of plants when it is advantageous to do so.
Technical Abstract: Promoter analyses can be compromised by interactions between the test promoter and those driving the expression of other genes within the same construct. Our laboratory is engaged in identifying and characterizing promoters that will be useful in controlling transgene expression in monocot crops. For this purpose, we have built the pGPro1 binary vector construct, a pGreen derived binary vector that contains a useful multiple cloning region allowing the creation of transcriptional or translational fusions to a gusA-enhanced Green Fluorescent Protein bifunctional reporter gene. The pGPro1 vector has many useful features that make it an efficient tool for Agrobacterium-mediated transformation and most importantly, its design minimizes the potential impact the promoter/selectable marker gene cassette has on the fidelity of the expression controlled by the test promoter. We demonstrate that the pGPro1 T-DNA is functional in both transient expression assays in onion cells and transgenic rice plants. This new binary vector will be a useful tool for characterizing promoter function in transgenic monocot plants.