Use of real-time RT-PCR for the detection of allelic expression of an imprinted gene

  • Authors:
    • Tetsuji Suda
    • Motonobu Katoh
    • Masaharu Hiratsuka
    • Mutsunori Fujiwara
    • Yoshito Irizawa
    • Mitsuo Oshimura
  • View Affiliations

  • Published online on: August 1, 2003     https://doi.org/10.3892/ijmm.12.2.243
  • Pages: 243-246
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Abstract

Measurement of the relative amounts of transcripts from two alleles is important in the study of imprinted genes, since quantitative differences that vary among tissues or individuals, and subtle differences in the ratio of allelic expression can have pathobiological significance. Discrimination of alleles is commonly based on PCR, followed by restriction endonuclease digestion to recognize a polymorphic site. However, the use of restriction enzymes misses most of the available single nucleotide polymorphisms. Practically, it requires substantial post-PCR analyses including the restriction enzyme digestion and gel electrophoresis, all of which increase turn around time. Taking advantage of our previous study identifying lung adenocarcinomas displaying biallelic expression of the imprinted gene MEST, we investigated the validity of a method of allelic discrimination in a real-time PCR assay using allele-specific probes. Allelic expression of the MEST gene in the range of 4-fold differences was detected. This new method should enhance our ability to rapidly and accurately assess allelic expression of imprinted genes in a number of samples.

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August 2003
Volume 12 Issue 2

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Suda T, Katoh M, Hiratsuka M, Fujiwara M, Irizawa Y and Oshimura M: Use of real-time RT-PCR for the detection of allelic expression of an imprinted gene. Int J Mol Med 12: 243-246, 2003
APA
Suda, T., Katoh, M., Hiratsuka, M., Fujiwara, M., Irizawa, Y., & Oshimura, M. (2003). Use of real-time RT-PCR for the detection of allelic expression of an imprinted gene. International Journal of Molecular Medicine, 12, 243-246. https://doi.org/10.3892/ijmm.12.2.243
MLA
Suda, T., Katoh, M., Hiratsuka, M., Fujiwara, M., Irizawa, Y., Oshimura, M."Use of real-time RT-PCR for the detection of allelic expression of an imprinted gene". International Journal of Molecular Medicine 12.2 (2003): 243-246.
Chicago
Suda, T., Katoh, M., Hiratsuka, M., Fujiwara, M., Irizawa, Y., Oshimura, M."Use of real-time RT-PCR for the detection of allelic expression of an imprinted gene". International Journal of Molecular Medicine 12, no. 2 (2003): 243-246. https://doi.org/10.3892/ijmm.12.2.243