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

162 Scopus citations

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 - Nov 1996

ASJC Scopus subject areas

  • Physiology
  • Genetics
  • Plant Science

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    Pang, S. Z., DeBoer, D. L., Wan, Y., Ye, G., Layton, J. G., Neher, M. K., Armstrong, C. L., Fry, J. E., Hinchee, M. A. W., & 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