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Plant Molecular Biology Reporter

, Volume 19, Issue 1, pp 13–26 | Cite as

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

  • S. Luke Mankin
  • William F. Thompson
Genetic Resources

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.

Key words

ER-localized GFP intron reporter gene toxicity transformation 

Abbreviations

35SP

caulflower mosaic virus 35S promoter

CaMV

cauliflower mosaic virus

GFP

green fluorescent protein

gusINT

β-glucuronidase cDNA with the PIV2 intron

lucINT

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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Copyright information

© International Society for Plant Molecular Biology 2001

Authors and Affiliations

  1. 1.Department of BotanyNC State UniversityRaleighUSA
  2. 2.Departments of Botany, Genetics, and Crop ScienceNC State UniversityRaleighUSA

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