An improved green fluorescent protein gene as a vital marker in plants

Sheng Zhi Pang, David L. DeBoer, Yuechun Wan, Guangning Ye, Jeanne G. Layton, Margaret K. Neher, Charles L. Armstrong, Joyce E. Fry, Maud A.W. Hinchee, Michael E Fromm

Research output: Contribution to journalArticle

160 Citations (Scopus)

Abstract

A synthetic green fluorescent protein (GFP) gene (pgfp) was constructed to improve GFP expression in plants. Corn and tobacco protoplast transient assays showed that pgfp gave about 20-fold brighter fluorescence than the wild-type gene (gfp). Replacement of the serine at position 65 with a threonine (S65Tpgfp) or a cysteine (S65Cpgfp) yielded 100- to 120-fold brighter fluorescence than wild-type gfp upon excitation with 490-nm light. Incorporation of a plant intron into the coding region yielded an additional 1.4-fold improvement, for a cumulative improvement of about 150-fold in fluorescence at 490-nm excitation. Various versions of pgfp were also stably introduced into corn, wheat, tobacco, and Arabidopsis plants. Bright-green fluorescence was observed with a fluorescence microscope in virtually all examined tissues of transgenic monocots and dicots. In the case of Arabidopsis, expression of the pgfp gene under the enhanced 35S promoter of the cauliflower mosaic virus produced green fluorescence that was readily detectable by eye using a hand-held, long-wave ultraviolet lamp and/or a black-light source.

Original languageEnglish (US)
Pages (from-to)893-900
Number of pages8
JournalPlant Physiology
Volume112
Issue number3
DOIs
StatePublished - Jan 1 1996

Fingerprint

Green Fluorescent Proteins
green fluorescent protein
Fluorescence
fluorescence
Genes
genes
Arabidopsis
Zea mays
Tobacco
tobacco
Caulimovirus
Light
corn
Protoplasts
Cauliflower mosaic virus
Threonine
Magnoliopsida
Liliopsida
threonine
microscopes

ASJC Scopus subject areas

  • Physiology
  • Genetics
  • Plant Science

Cite this

Pang, S. Z., DeBoer, D. L., Wan, Y., Ye, G., Layton, J. G., Neher, M. K., ... Fromm, M. E. (1996). An improved green fluorescent protein gene as a vital marker in plants. Plant Physiology, 112(3), 893-900. https://doi.org/10.1104/pp.112.3.893

An improved green fluorescent protein gene as a vital marker in plants. / Pang, Sheng Zhi; DeBoer, David L.; Wan, Yuechun; Ye, Guangning; Layton, Jeanne G.; Neher, Margaret K.; Armstrong, Charles L.; Fry, Joyce E.; Hinchee, Maud A.W.; Fromm, Michael E.

In: Plant Physiology, Vol. 112, No. 3, 01.01.1996, p. 893-900.

Research output: Contribution to journalArticle

Pang, SZ, DeBoer, DL, Wan, Y, Ye, G, Layton, JG, Neher, MK, Armstrong, CL, Fry, JE, Hinchee, MAW & Fromm, ME 1996, 'An improved green fluorescent protein gene as a vital marker in plants', Plant Physiology, vol. 112, no. 3, pp. 893-900. https://doi.org/10.1104/pp.112.3.893
Pang SZ, DeBoer DL, Wan Y, Ye G, Layton JG, Neher MK et al. An improved green fluorescent protein gene as a vital marker in plants. Plant Physiology. 1996 Jan 1;112(3):893-900. https://doi.org/10.1104/pp.112.3.893
Pang, Sheng Zhi ; DeBoer, David L. ; Wan, Yuechun ; Ye, Guangning ; Layton, Jeanne G. ; Neher, Margaret K. ; Armstrong, Charles L. ; Fry, Joyce E. ; Hinchee, Maud A.W. ; Fromm, Michael E. / An improved green fluorescent protein gene as a vital marker in plants. In: Plant Physiology. 1996 ; Vol. 112, No. 3. pp. 893-900.
@article{c28985bee1ce410fa01184efbe3f112f,
title = "An improved green fluorescent protein gene as a vital marker in plants",
abstract = "A synthetic green fluorescent protein (GFP) gene (pgfp) was constructed to improve GFP expression in plants. Corn and tobacco protoplast transient assays showed that pgfp gave about 20-fold brighter fluorescence than the wild-type gene (gfp). Replacement of the serine at position 65 with a threonine (S65Tpgfp) or a cysteine (S65Cpgfp) yielded 100- to 120-fold brighter fluorescence than wild-type gfp upon excitation with 490-nm light. Incorporation of a plant intron into the coding region yielded an additional 1.4-fold improvement, for a cumulative improvement of about 150-fold in fluorescence at 490-nm excitation. Various versions of pgfp were also stably introduced into corn, wheat, tobacco, and Arabidopsis plants. Bright-green fluorescence was observed with a fluorescence microscope in virtually all examined tissues of transgenic monocots and dicots. In the case of Arabidopsis, expression of the pgfp gene under the enhanced 35S promoter of the cauliflower mosaic virus produced green fluorescence that was readily detectable by eye using a hand-held, long-wave ultraviolet lamp and/or a black-light source.",
author = "Pang, {Sheng Zhi} and DeBoer, {David L.} and Yuechun Wan and Guangning Ye and Layton, {Jeanne G.} and Neher, {Margaret K.} and Armstrong, {Charles L.} and Fry, {Joyce E.} and Hinchee, {Maud A.W.} and Fromm, {Michael E}",
year = "1996",
month = "1",
day = "1",
doi = "10.1104/pp.112.3.893",
language = "English (US)",
volume = "112",
pages = "893--900",
journal = "Plant Physiology",
issn = "0032-0889",
publisher = "American Society of Plant Biologists",
number = "3",

}

TY - JOUR

T1 - An improved green fluorescent protein gene as a vital marker in plants

AU - Pang, Sheng Zhi

AU - DeBoer, David L.

AU - Wan, Yuechun

AU - Ye, Guangning

AU - Layton, Jeanne G.

AU - Neher, Margaret K.

AU - Armstrong, Charles L.

AU - Fry, Joyce E.

AU - Hinchee, Maud A.W.

AU - Fromm, Michael E

PY - 1996/1/1

Y1 - 1996/1/1

N2 - A synthetic green fluorescent protein (GFP) gene (pgfp) was constructed to improve GFP expression in plants. Corn and tobacco protoplast transient assays showed that pgfp gave about 20-fold brighter fluorescence than the wild-type gene (gfp). Replacement of the serine at position 65 with a threonine (S65Tpgfp) or a cysteine (S65Cpgfp) yielded 100- to 120-fold brighter fluorescence than wild-type gfp upon excitation with 490-nm light. Incorporation of a plant intron into the coding region yielded an additional 1.4-fold improvement, for a cumulative improvement of about 150-fold in fluorescence at 490-nm excitation. Various versions of pgfp were also stably introduced into corn, wheat, tobacco, and Arabidopsis plants. Bright-green fluorescence was observed with a fluorescence microscope in virtually all examined tissues of transgenic monocots and dicots. In the case of Arabidopsis, expression of the pgfp gene under the enhanced 35S promoter of the cauliflower mosaic virus produced green fluorescence that was readily detectable by eye using a hand-held, long-wave ultraviolet lamp and/or a black-light source.

AB - A synthetic green fluorescent protein (GFP) gene (pgfp) was constructed to improve GFP expression in plants. Corn and tobacco protoplast transient assays showed that pgfp gave about 20-fold brighter fluorescence than the wild-type gene (gfp). Replacement of the serine at position 65 with a threonine (S65Tpgfp) or a cysteine (S65Cpgfp) yielded 100- to 120-fold brighter fluorescence than wild-type gfp upon excitation with 490-nm light. Incorporation of a plant intron into the coding region yielded an additional 1.4-fold improvement, for a cumulative improvement of about 150-fold in fluorescence at 490-nm excitation. Various versions of pgfp were also stably introduced into corn, wheat, tobacco, and Arabidopsis plants. Bright-green fluorescence was observed with a fluorescence microscope in virtually all examined tissues of transgenic monocots and dicots. In the case of Arabidopsis, expression of the pgfp gene under the enhanced 35S promoter of the cauliflower mosaic virus produced green fluorescence that was readily detectable by eye using a hand-held, long-wave ultraviolet lamp and/or a black-light source.

UR - http://www.scopus.com/inward/record.url?scp=10544244586&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=10544244586&partnerID=8YFLogxK

U2 - 10.1104/pp.112.3.893

DO - 10.1104/pp.112.3.893

M3 - Article

VL - 112

SP - 893

EP - 900

JO - Plant Physiology

JF - Plant Physiology

SN - 0032-0889

IS - 3

ER -