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Acta Biologica Hungarica

, Volume 55, Issue 1–4, pp 157–161 | Cite as

Real-Time Quantitative RT-PCR Method for Estimation of mRNA Level of CCAAT/Enhancer Binding Protein in the Central Nervous System of Lymnaea Stagnalis

  • D. HatakeyamaEmail author
  • Hisayo Sadamoto
  • E. Ito
Article

Abstract

The fluorescence-based real-time reverse transcription polymerase chain reaction (RT-PCR) is becoming widely used to quantify mRNA level in cells and tissues and is now a. crucial tool for basic biological researches and biotechnology. In the present study, on the basis of the real-time quantitative RT-RCR, we detected and quantified mRNA copies of the transcription factor, CCAAT/enhancer binding protein (C/EBP; an immediate-early gene that is involved in synaptic plasticity and learning and memory) in the central nervous system of the pond snail Lymnaea stagnalis. We designed the primer set and the probe in the specific insert for the detection of Lymnaea C/EBP (LymC/EBP) clone 1. This insert is not contained in LymC/EBP clone 2 by alternative splicing. The copy number of LymC/EBP clone 1 was linearly decreased relative to the dilution of cDNA, and it was estimated 30 copies in test sample. The availability of the present study showed that the real-time quantitative RT-PCR technique is more accurate and more specific for the detection and quantification of the mRNA level of genes in L. stagnalis than the other PCR methods.

Keywords

C/EBP mRNA qRT-PCR CNS Lymnaea 

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Copyright information

© Akadémiai Kiadó, Budapest 2004

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  1. 1.Division of Biological SciencesGraduate School of ScienceSapporoJapan
  2. 2.Division of Innovative Research, Creative Research Initiative “Sousei” (CRIS)Hokkaido UniversitySapporoJapan

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