Submitted to: Plant Molecular Biology
Publication Type: Peer reviewed journal
Publication Acceptance Date: 3/22/2005
Publication Date: 5/1/2005
Citation: Shigaki, T., Vyzasatya, R.R., Sivitz, A.B., Ward, J.W., Sze, H., Hirschi, K.D. 2005. The Cre-loxP recombination-based reporter system for plant transcriptional expression studies. Plant Molecular Biology. 58(1):65-73. Interpretive Summary: Using molecular biology can be expensive and intimidating. For example, often cloning procedures require expensive enzymes and a wealth of knowledge. In this study, we have developed tools by which any lab can clone genes regardless of their economic status and prior experience. We have thus circumvented the problems associated with cloning by providing an inexpensive and easy procedure that should become a general resource to the scientific community.
Technical Abstract: To facilitate the characterization of plant genes, the Cre-loxP site-specific recombination system was adapted to make reporter vectors for plant expression studies. This system allows promoter fragments to be cloned into a small vector (univector) and subsequently recombined in vitro with binary vectors containing different reporter genes precisely at near-perfect efficiency. We have constructed univector-adapted vectors with three reporters, beta-glucuronidase, luciferase, and green fluorescent protein, and a BASTA-resistance gene for selection of plant transformants. Expression in plants using the new system was validated by comparison to conventional reporter vectors. These new vectors are efficient and economical alternatives to the other plant reporter vectors currently available. The royalty-free Cre-loxP system serves as a platform for the future expansion of recombination-based cloning vectors for plant research.