Research Article | Published:

Green fluorescent protein as a marker for expression of a second gene in transgenic plants

Nature Biotechnology volume 17, pages 11251129 (1999) | Download Citation

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Abstract

The use of transgenic crops has generated concerns about transgene movement to unintended hosts and the associated ecological consequences. Moreover, the in-field monitoring of transgene expression is of practical concern (e.g., the underexpression of an herbicide tolerance gene in crop plants that are due to be sprayed with herbicide). A solution to these potential problems is to monitor the presence and expression of an agronomically important gene by linking it to a marker gene, such as GFP. Here we show that GFP fluorescence can indicate expression of the Bacillus thuringiensus cry1Ac gene when co-introduced into tobacco and oilseed rape, as demonstrated by insect bioassays and western blot analysis. Furthermore we conducted two seasons of field experiments to characterize the performance of three different GFP genes in transgenic tobacco. The best gene tested was mGFP5er, a mutagenized GFP gene that is targeted to the endoplasmic reticulum. We also demonstrated that host plants synthesizing GFP in the field suffered no fitness costs.

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Acknowledgements

We thank Jim Haseloff for the gift of the mGFP4 and mGFP5er constructs. We thank Jen Sheen and C.S. Prakash for the gift of the sGFP gene. We thank Guy Cardineau and Dow AgroSciences for the gift of the Bt cry1Ac gene, protein, and antisera. Research was funded by the USDA Biotechnology Risk Assessment grant 98-33522-6797, and NSF grant 9604528. We also thank Jimmy Meadows and Joe French at the Upper Piedmont Expression Research Station, as well as Sheila Branch, Tanya Chalk, Dean Chamberlain, Richard Emanuel, Mark Hens, Laura Hudson, Kevin Markham, Reggie Millwood, Alana Riddick, Shana Stovall, Elizabeth Tomlin, and Elyse Williams, for help in both the field and the lab.

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Affiliations

  1. Department of Biology, University of North Carolina, Greensboro, NC 27402-6174.

    • Brian K. Harper
    • , Stephen A. Mabon
    • , Staci M. Leffel
    • , Matthew D. Halfhill
    • , Harold A. Richards
    • , Kari A. Moyer
    •  & C. Neal Stewart Jr.

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Correspondence to C. Neal Stewart Jr.

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DOI

https://doi.org/10.1038/15114