Abstract
It makes little sense to invest time and effort in assaying biological activities of transgenic plants if the plants do not contain a functional gene of interest. Although this appears to be self-evident, many investigators attempt to use biological assays for the presence of the gene of interest prior to conducting informative molecular characterization of transgenic plants. Where facile tests are available, such as those for β-glucuronidase (GUS) activity, this may be feasible. However, low levels of activity frequently make simple biochemical screening approaches unreliable. Of the several analytical techniques available, we have chosen to describe methods for polymerase chain reaction (PCR) and genomic DNA blot analysis: methods that are fundamental for confirming and characterizing gene insertion.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Southern, E. (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98, 503–511.
Taylor, B. H., Manhart, J. R., and Amasino, R. M. (1993) Isolation and characterization of plant DNAs, in Methods in Plant Molecular Biology and Biotechnology (Glick, B. R. and Thompson, J. E., eds.), CRC, Boca Raton, pp. 360.
Agudo, L. d. C, Gavidia, I., Perez-Bermudez, P., and Sugura, J. (1995) PEG precipitation, a required step for PCRF amplification of DNA from wild plants of Digitalis obscura L. Biotechniques 18, 766–768.
Xu, Y., Buchholz, W. G, DeRose, R. T, and Hall, T. C. (1995) Characterization of a rice gene family encoding root-specific proteins. Plant Mol. Biol. 27, 237–248.
Matzke, M. A. and Matzke, A. J. M. (1995) How and why do plants inactivate homologous (trans)genes? Plant Physiol. 107, 679–685.
Finnegan, J. and McElroy, D. (1994) Transgene inactivation: plants fight back! Biotechnology 12, 883–888.
Flavell, R. B. (1994) Inactivation of gene expression in plants as a consequence of specific sequence duplication. Proc. Natl. Acad. Sci. USA 91, 3490–3496.
Chamberlin, M. A. and Bowen, B. (1995) Nuclear changes in maize cells transformed by DNA-coated microprojectiles, in 24th Annual Keystone Symposia, Journal of Cellular Biochemistry, Hilton Head Island, SC, pp. 138.
Hall, T. C., Xu, Y., Huntley, C. C., Yu, H., Seay, J., Connell, J. C., Lepetit, M., Dong, J., Wallace, D., Way, M. O., and Buchholz, W. G. (1993) Transgene expression and agronomic improvement of rice. Phil. Trans. R. Soc. Lond. B 342, 197–201.
Chalfie, M., Tu, Y., Euskirchen, G., Ward, W. W., and Prasher, D. G. (1994) Green fluorescent protein as a marker for gene expression. Science 263, 802–805.
Xu, Y., Harris-Haller, L. W., McCollum, J. C., Hardin, S. H., and Hall, T. C. (1993) Nuclear gene encoding cytosolic triosephosphate isomerase from rice (Oryza sativa L.). Plant Physiol. 102, 697.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1998 Humana Press Inc.
About this protocol
Cite this protocol
Buchholz, W.G., Connell, J.P., Kumpatla, S.P., Hall, T.C. (1998). Molecular Analysis of Transgenic Rice. In: Foster, G.D., Taylor, S.C. (eds) Plant Virology Protocols. Methods in Molecular Biology™, vol 81. Humana Press. https://doi.org/10.1385/0-89603-385-6:397
Download citation
DOI: https://doi.org/10.1385/0-89603-385-6:397
Publisher Name: Humana Press
Print ISBN: 978-0-89603-385-6
Online ISBN: 978-1-59259-566-2
eBook Packages: Springer Protocols