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. 1990 Dec;87(23):9183–9187. doi: 10.1073/pnas.87.23.9183

Variable patterns of expression of luciferase in transgenic tobacco leaves.

W M Barnes 1
PMCID: PMC55128  PMID: 2251262

Abstract

A carboxyl-terminally modified firefly luciferase, encoded as a gene fusion to the neomycin phosphotransferase gene (which confers kanamycin resistance), was found to be enzymatically active for both enzymes when expressed in bacteria and in transgenic plants. A military-type starlight vision system was used to conveniently analyze the pattern of gene expression in transgenic tobacco plant leaves. Transgenic tobacco plants which expressed luciferase uniformly in all areas of the leaf, and assays for luciferin, demonstrated that luciferin rapidly penetrates all regions of a tobacco leaf in at least two dimensions. Depending on the test gene structure or, presumably, on the transferred DNA (T-DNA) insertional context, other transgenic plants were obtained that expressed luciferase with a wide range of nonuniform patterns from nominally the same cauliflower mosaic virus 35S promoter. For instance, the veins can be dark, while only the interveinal regions of the leaf lamina glow, or only the small capillary veins glow, or only the major veins glow. Local and/or systemic induction in response to wounding was also demonstrated.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. A simple and general method for transferring genes into plants. Science. 1985 Mar 8;227(4691):1229–1231. doi: 10.1126/science.227.4691.1229. [DOI] [PubMed] [Google Scholar]
  2. Abel P. P., Nelson R. S., De B., Hoffmann N., Rogers S. G., Fraley R. T., Beachy R. N. Delay of disease development in transgenic plants that express the tobacco mosaic virus coat protein gene. Science. 1986 May 9;232(4751):738–743. doi: 10.1126/science.3457472. [DOI] [PubMed] [Google Scholar]
  3. An G., Mitra A., Choi H. K., Costa M. A., An K., Thornburg R. W., Ryan C. A. Functional analysis of the 3' control region of the potato wound-inducible proteinase inhibitor II gene. Plant Cell. 1989 Jan;1(1):115–122. doi: 10.1105/tpc.1.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beck E., Ludwig G., Auerswald E. A., Reiss B., Schaller H. Nucleotide sequence and exact localization of the neomycin phosphotransferase gene from transposon Tn5. Gene. 1982 Oct;19(3):327–336. doi: 10.1016/0378-1119(82)90023-3. [DOI] [PubMed] [Google Scholar]
  5. Benfey P. N., Chua N. H. Regulated genes in transgenic plants. Science. 1989 Apr 14;244(4901):174–181. doi: 10.1126/science.244.4901.174. [DOI] [PubMed] [Google Scholar]
  6. Bevan M., Barnes W. M., Chilton M. D. Structure and transcription of the nopaline synthase gene region of T-DNA. Nucleic Acids Res. 1983 Jan 25;11(2):369–385. doi: 10.1093/nar/11.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bishop P. D., Pearce G., Bryant J. E., Ryan C. A. Isolation and characterization of the proteinase inhibitor-inducing factor from tomato leaves. Identity and activity of poly- and oligogalacturonide fragments. J Biol Chem. 1984 Nov 10;259(21):13172–13177. [PubMed] [Google Scholar]
  8. Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ellenson J. L., Raba R. M. Gas Exchange and Phytoluminography of Single Red Kidney Bean Leaves during Periods of Induced Stomatal Oscillations: A Demonstration of an Integrated, Spatially Resolving Physiometric Technique. Plant Physiol. 1983 May;72(1):90–95. doi: 10.1104/pp.72.1.90. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gardner R. C., Howarth A. J., Hahn P., Brown-Luedi M., Shepherd R. J., Messing J. The complete nucleotide sequence of an infectious clone of cauliflower mosaic virus by M13mp7 shotgun sequencing. Nucleic Acids Res. 1981 Jun 25;9(12):2871–2888. doi: 10.1093/nar/9.12.2871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gould S. J., Keller G. A., Hosken N., Wilkinson J., Subramani S. A conserved tripeptide sorts proteins to peroxisomes. J Cell Biol. 1989 May;108(5):1657–1664. doi: 10.1083/jcb.108.5.1657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. KELLENBERGER E., LARK K. G., BOLLE A. Amino acid dependent control of DNA synthesis in bacteria and vegetative phage. Proc Natl Acad Sci U S A. 1962 Oct 15;48:1860–1868. doi: 10.1073/pnas.48.10.1860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lee J. S., Brown W. E., Graham J. S., Pearce G., Fox E. A., Dreher T. W., Ahern K. G., Pearson G. D., Ryan C. A. Molecular characterization and phylogenetic studies of a wound-inducible proteinase inhibitor I gene in Lycopersicon species. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7277–7281. doi: 10.1073/pnas.83.19.7277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ow D. W., DE Wet J. R., Helinski D. R., Howell S. H., Wood K. V., Deluca M. Transient and stable expression of the firefly luciferase gene in plant cells and transgenic plants. Science. 1986 Nov 14;234(4778):856–859. doi: 10.1126/science.234.4778.856. [DOI] [PubMed] [Google Scholar]
  15. STREHLER B. L., ARNOLD W. Light production by green plants. J Gen Physiol. 1951 Jul;34(6):809–820. doi: 10.1085/jgp.34.6.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  17. Sciaky D., Montoya A. L., Chilton M. D. Fingerprints of Agrobacterium Ti plasmids. Plasmid. 1978 Feb;1(2):238–253. doi: 10.1016/0147-619x(78)90042-2. [DOI] [PubMed] [Google Scholar]
  18. Shah D. M., Horsch R. B., Klee H. J., Kishore G. M., Winter J. A., Tumer N. E., Hironaka C. M., Sanders P. R., Gasser C. S., Aykent S., Siegel N. R., Rogers S. G., Fraley R. T. Engineering herbicide tolerance in transgenic plants. Science. 1986 Jul 25;233(4762):478–481. doi: 10.1126/science.233.4762.478. [DOI] [PubMed] [Google Scholar]
  19. Teeri T. H., Lehväslaiho H., Franck M., Uotila J., Heino P., Palva E. T., Van Montagu M., Herrera-Estrella L. Gene fusions to lacZ reveal new expression patterns of chimeric genes in transgenic plants. EMBO J. 1989 Feb;8(2):343–350. doi: 10.1002/j.1460-2075.1989.tb03383.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Thornburg R. W., An G., Cleveland T. E., Johnson R., Ryan C. A. Wound-inducible expression of a potato inhibitor II-chloramphenicol acetyltransferase gene fusion in transgenic tobacco plants. Proc Natl Acad Sci U S A. 1987 Feb;84(3):744–748. doi: 10.1073/pnas.84.3.744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. de Wet J. R., Wood K. V., DeLuca M., Helinski D. R., Subramani S. Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol. 1987 Feb;7(2):725–737. doi: 10.1128/mcb.7.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]

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