The Cre-loxP recombination-based reporter system for plant transcriptional expression studies

Toshiro Shigaki; Ravindranadha R. Vyzasatya; Ali B. Sivitz; John M. Ward; Heven Sze; Kendal D. Hirschi

doi:10.1007/s11103-005-4323-3

The Cre-loxP recombination-based reporter system for plant transcriptional expression studies

Toshiro Shigaki, Ravindranadha R. Vyzasatya, Ali B. Sivitz, John M. Ward, Heven Sze, Kendal D. Hirschi

Administration (CBS)

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

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, β-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.

Original language	English (US)
Pages (from-to)	65-73
Number of pages	9
Journal	Plant molecular biology
Volume	58
Issue number	1
DOIs	https://doi.org/10.1007/s11103-005-4323-3
State	Published - May 2005

Bibliographical note

Funding Information:
We thank Stephen J. Elledge, Bonnie Bartel, and Sherry LeClere for the template vectors. We extend our thanks to Jay Morris for technical assistance in luciferase imaging, and Ian Rees for artwork. This study was funded by the National Science Foundation 0209777 to K.H., 0209792 to J.W., and 0209788 to H.S.

Keywords

Cre-loxP recombination
GFP
GUS
Gene expression
Luciferase
Promoter

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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, β-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.",

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AU - Ward, John M.

AU - Sze, Heven

AU - Hirschi, Kendal D.

N1 - Funding Information: We thank Stephen J. Elledge, Bonnie Bartel, and Sherry LeClere for the template vectors. We extend our thanks to Jay Morris for technical assistance in luciferase imaging, and Ian Rees for artwork. This study was funded by the National Science Foundation 0209777 to K.H., 0209792 to J.W., and 0209788 to H.S.

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The Cre-loxP recombination-based reporter system for plant transcriptional expression studies

Abstract

Bibliographical note

Keywords

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