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Gene fusions to lacZ reveal new expression patterns of chimeric genes in transgenic plants

TH Teeri, H Lehväslaiho, M Franck, J Uotila, P Heino, ET Palva, Marc Van Montagu UGent and L Herrera-Estrella (1989) EMBO JOURNAL. 8(2). p.343-350
abstract
The lacZ gene of Escherichia coli, coding for beta-galactosidase, is a widely used reporter gene for gene expression studies in microbial and animal systems. To demonstrate that it is also a powerful reporter gene in plants, lacZ was fused to 5' regulatory elements of several genes known to be functional in plant cells. By measuring LacZ activities in transgenic plants containing these gene constructs, we showed that the reporter is correctly monitoring the regulatory properties of the well-characterized promoters fused to lacZ. beta-Galactosidase was assayed directly in plant extracts when they contained high levels of LacZ or, when LacZ was expressed at low level, by separating the endogenous and LacZ activities electrophoretically and detecting the enzymes with a fluorogenic substrate. The most outstanding property of the marker is its amenability to histochemical detection. Due to its stability, LacZ can be fixed in the tissue with glutaraldehyde without loss of activity and detected with high resolution by using XGal. We could reveal expression patterns unnoticed earlier for many of the regulatory elements studied. The chlorophyll a/b binding protein gene, expressed at very high levels in green tissues, is also expressed at a low level in the vascular cylinder of the root. The Agrobacterium T-DNA gene encoding octopine synthase is especially active in the epidermis of the root tip and the TR2' gene was shown to be root specific in the intact plant and stimulated by wounding in the leaf tissue. The TR1' gene, fused to nptII, shows similar characteristics suggesting co-regulation of this tightly linked dual promoter.
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author
organization
year
type
journalArticle (original)
publication status
published
subject
journal title
EMBO JOURNAL
Embo J.
volume
8
issue
2
pages
343 - 350
Web of Science type
Article
ISSN
0261-4189
language
English
UGent publication?
yes
classification
A1
copyright statement
I don't know the status of the copyright for this publication
id
1910253
handle
http://hdl.handle.net/1854/LU-1910253
alternative location
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC400812/?tool=pubmed
date created
2011-09-28 11:02:07
date last changed
2016-12-19 15:46:25
@article{1910253,
  abstract     = {The lacZ gene of Escherichia coli, coding for beta-galactosidase, is a widely used reporter gene for gene expression studies in microbial and animal systems. To demonstrate that it is also a powerful reporter gene in plants, lacZ was fused to 5' regulatory elements of several genes known to be functional in plant cells. By measuring LacZ activities in transgenic plants containing these gene constructs, we showed that the reporter is correctly monitoring the regulatory properties of the well-characterized promoters fused to lacZ. beta-Galactosidase was assayed directly in plant extracts when they contained high levels of LacZ or, when LacZ was expressed at low level, by separating the endogenous and LacZ activities electrophoretically and detecting the enzymes with a fluorogenic substrate. The most outstanding property of the marker is its amenability to histochemical detection. Due to its stability, LacZ can be fixed in the tissue with glutaraldehyde without loss of activity and detected with high resolution by using XGal. We could reveal expression patterns unnoticed earlier for many of the regulatory elements studied. The chlorophyll a/b binding protein gene, expressed at very high levels in green tissues, is also expressed at a low level in the vascular cylinder of the root. The Agrobacterium T-DNA gene encoding octopine synthase is especially active in the epidermis of the root tip and the TR2' gene was shown to be root specific in the intact plant and stimulated by wounding in the leaf tissue. The TR1' gene, fused to nptII, shows similar characteristics suggesting co-regulation of this tightly linked dual promoter.},
  author       = {Teeri, TH and Lehv{\"a}slaiho, H and Franck, M and Uotila, J and Heino, P and Palva, ET and Van Montagu, Marc and Herrera-Estrella, L},
  issn         = {0261-4189},
  journal      = {EMBO JOURNAL},
  language     = {eng},
  number       = {2},
  pages        = {343--350},
  title        = {Gene fusions to lacZ reveal new expression patterns of chimeric genes in transgenic plants},
  url          = {http://www.ncbi.nlm.nih.gov/pmc/articles/PMC400812/?tool=pubmed},
  volume       = {8},
  year         = {1989},
}

Chicago
Teeri, TH, H Lehväslaiho, M Franck, J Uotila, P Heino, ET Palva, Marc Van Montagu, and L Herrera-Estrella. 1989. “Gene Fusions to lacZ Reveal New Expression Patterns of Chimeric Genes in Transgenic Plants.” Embo Journal 8 (2): 343–350.
APA
Teeri, T., Lehväslaiho, H., Franck, M., Uotila, J., Heino, P., Palva, E., Van Montagu, M., et al. (1989). Gene fusions to lacZ reveal new expression patterns of chimeric genes in transgenic plants. EMBO JOURNAL, 8(2), 343–350.
Vancouver
1.
Teeri T, Lehväslaiho H, Franck M, Uotila J, Heino P, Palva E, et al. Gene fusions to lacZ reveal new expression patterns of chimeric genes in transgenic plants. EMBO JOURNAL. 1989;8(2):343–50.
MLA
Teeri, TH, H Lehväslaiho, M Franck, et al. “Gene Fusions to lacZ Reveal New Expression Patterns of Chimeric Genes in Transgenic Plants.” EMBO JOURNAL 8.2 (1989): 343–350. Print.