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

Research output: Contribution to journalArticlepeer-review

7 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 languageEnglish (US)
Pages (from-to)65-73
Number of pages9
JournalPlant molecular biology
Volume58
Issue number1
DOIs
StatePublished - 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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