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New green fluorescent protein genes for plant transformation: Intron-containing, ER-localized, and soluble-modified

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Abstract

We describe several new modifications of theAequorea victoria green fluorescent protein (GFP) gene. TheerGFP5 INTreporter gene combines the PIV2 intron fromgus INTandLUC INTwith the ER-localizedmGFP4-ER gene. TheerGFP6 INT, erGFP7INT, anderBFP8 INTgenes also include the fluorophore and solubility modifications of smGFP, smRS-GFP, and smBFP, respectively. A parallel set of reporter genes (erGFP5, erGFP6, erGFP7, anderBFP8) is otherwise identical to the respectiveerGFP INTgenes but lacks the PIV2 intron. The intron-containing genes are expressed in plant cells but not in bacteria, allowing detection of plant cell expression in the presence ofAgrobacterium during the early stages of transformation. Transient expression of theerGFP INTanderGFP genes is comparable in tobacco and maize suspension culture protoplasts, indicating that the PIV2 intron is spliced effectively in both monocotyledonous and dicotyledonous plant species.

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Abbreviations

35SP:

caulflower mosaic virus 35S promoter

CaMV:

cauliflower mosaic virus

GFP:

green fluorescent protein

gus INT :

β-glucuronidase cDNA with the PIV2 intron

luc INT :

luciferase cDNA with the PIV2 intron

nosT:

nopaline synthase 3é region

nptII:

neomycin phosphotransferase II

PIV2:

synthetic intron derived from the second intron of the potatoST-LS1 gene

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Author notes

  1. S. Luke Mankin

    Present address: BASF Plant Sciences LLC, 26 Davis Drive, P.O. Box 13528, 27709-3528, Research Triangle Park, NC, USA

Authors and Affiliations

  1. Department of Botany, NC State University, 27695-7612, Raleigh, NC, USA

    S. Luke Mankin

  2. Departments of Botany, Genetics, and Crop Science, NC State University, 27695-7612, Raleigh, NC, USA

    William F. Thompson

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  2. William F. Thompson
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Correspondence to S. Luke Mankin.

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Luke Mankin, S., Thompson, W.F. New green fluorescent protein genes for plant transformation: Intron-containing, ER-localized, and soluble-modified. Plant Mol Biol Rep 19, 13–26 (2001). https://doi.org/10.1007/BF02824074

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