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Transgenic Plants: Methods and Protocols pp 291–311Cite as

Analysis of Gene Expression in Transgenic Plants

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 286))

Summary

Recent years have seen a huge increase and improvement in techniques for analysis of transgene expression in plants. The analysis of RNA frequently provides a vital link between changes in enzyme levels and/or metabolites and the phenotype. This chapter focuses on RNA-based techniques for the analysis of transgene expression, beginning with the extraction of RNA and its evaluation in terms of purity and integrity by spectrophotometry and gel electrophoresis, respectively. Common methods of transcript analysis by Northern and dot-blot hybridizations using nonradioactive probing methods are described. A protocol for reverse-transcriptase polymerase chain reaction (RT-PCR) as a method of establishing transgene expression qualitatively, as well as a procedure for quantitative RT-PCR for comparing relative abundance of transcript levels of two or more genes are described. Lastly, a protocol for localization of RNA transcripts within tissues by in situ hybridization is included.

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Author information

Authors and Affiliations

  1. Department of Plant Biology, University of New Hampshire, Durham, NH

    Andrew F. Page & Subhash C. Minocha

Authors
  1. Andrew F. Page
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  2. Subhash C. Minocha
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Editor information

Editors and Affiliations

  1. Instituto Valenciano de Investigaciones Agrarias, Valencia, Spain

    Leandro Peña

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© 2005 Humana Press Inc.

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Page, A.F., Minocha, S.C. (2005). Analysis of Gene Expression in Transgenic Plants. In: Peña, L. (eds) Transgenic Plants: Methods and Protocols. Methods in Molecular Biology™, vol 286. Humana Press. https://doi.org/10.1385/1-59259-827-7:291

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  • DOI: https://doi.org/10.1385/1-59259-827-7:291

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-263-6

  • Online ISBN: 978-1-59259-827-4

  • eBook Packages: Springer Protocols

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