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A new method for analyzing gene expression based on frequency analysis of RT-PCR products obtained with degenerate primers

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Abstract

This paper presents a new method of gene expression analysis: frequency analysis of RT-PCR products obtained with degenerate primers (FAPP). The main advantage of the new approach compared to the present methods of gene expression analysis is that it is applicable to non-model plant objects whose gene sequences are unknown. The advantages and disadvantages of FAPP are described in detail using data on calcium-dependent protein kinase, stilbene synthase, and phenylalanine ammonia-lyase gene expression in two cell cultures of Vitis amurensis. We compared the expression profiles obtained by FAPP to those obtained by real-time PCR and expressed sequence tags.

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Abbreviations

CDPK:

Calcium-dependent protein kinase

FAPP:

Frequency analysis of RT-PCR products obtained with degenerate primers

EST:

Expressed sequence tags

PAL:

Phenylalanine ammonia-lyase

STS:

Stilbene synthase

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Acknowledgments

This work was supported by a grant from the President of Russia (MК-714.2008.4), by grants of the Far East Division of the Russian Academy of Sciences (06-III-A-06-146) and a grant from the Russian Science Support Foundation.

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Authors and Affiliations

  1. Laboratory of Biotechnology, Institute of Biology and Soil Science, Far East Branch of Russian Academy of Sciences, Stoletija Str. 159, 690022, Vladivostok, Russia

    Konstantin V. Kiselev & Alexandra S. Dubrovina

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  1. Konstantin V. Kiselev
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  2. Alexandra S. Dubrovina
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Correspondence to Konstantin V. Kiselev.

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Communicated by L. A. Kleczkowski.

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Kiselev, K.V., Dubrovina, A.S. A new method for analyzing gene expression based on frequency analysis of RT-PCR products obtained with degenerate primers. Acta Physiol Plant 32, 495–502 (2010). https://doi.org/10.1007/s11738-009-0426-9

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  • DOI: https://doi.org/10.1007/s11738-009-0426-9

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