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A multiplex RT‐PCR assay for analysis of relative transcript levels of different members of multigene families: Application to Arabidopsis calmodulin gene family

A multiplex RT‐PCR assay for analysis of relative transcript levels of different members of... The high degree of conservation of nucleotide sequences among different members of a multigene family poses problems in analysis of expression patterns governed by each member of the gene family. In this report we describe a simple, semi‐quantitative and single tube multiplex RT‐PCR assay for simultaneous and relative expression analysis with an application to all the six members of Arabidopsis calmodulin multigene family. In the multiplex primer set, individual gene specific primers were derived from 3′‐untranslated region of the genes and a single common primer from the conserved exonic region. Transcriptional activation of all the members of the calmodulin gene family in response to touch was monitored. The results demonstrate that two of the genes are not regulated by touch; however, the other four that are induced by touch show a differential response including their kinetics of induction. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png IUBMB Life Wiley

A multiplex RT‐PCR assay for analysis of relative transcript levels of different members of multigene families: Application to Arabidopsis calmodulin gene family

IUBMB Life , Volume 46 (4) – Nov 1, 1998

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Publisher
Wiley Subscription Services, Inc., A Wiley Company
Copyright
Copyright © 1998 International Union of Biochemistry and Molecular Biology
ISSN
1521-6543
eISSN
1521-6551
DOI
10.1080/15216549800204232
Publisher site
See Article on Publisher Site

Abstract

The high degree of conservation of nucleotide sequences among different members of a multigene family poses problems in analysis of expression patterns governed by each member of the gene family. In this report we describe a simple, semi‐quantitative and single tube multiplex RT‐PCR assay for simultaneous and relative expression analysis with an application to all the six members of Arabidopsis calmodulin multigene family. In the multiplex primer set, individual gene specific primers were derived from 3′‐untranslated region of the genes and a single common primer from the conserved exonic region. Transcriptional activation of all the members of the calmodulin gene family in response to touch was monitored. The results demonstrate that two of the genes are not regulated by touch; however, the other four that are induced by touch show a differential response including their kinetics of induction.

Journal

IUBMB LifeWiley

Published: Nov 1, 1998

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