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. Hatakeyama, Hisayo Sadamoto, Etsuro Ito

Research output: Contribution to journalArticle

13 Citations (Scopus)

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/μl 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.

Original languageEnglish
Pages (from-to)157-161
Number of pages5
JournalActa Biologica Hungarica
Volume55
Issue number1-4
DOIs
Publication statusPublished - 2004
Externally publishedYes

Fingerprint

CCAAT-Enhancer-Binding Proteins
Lymnaea
Lymnaea stagnalis
Polymerase chain reaction
Neurology
Transcription
nervous system
polymerase chain reaction
Reverse Transcription
central nervous system
binding proteins
clone
quantitative polymerase chain reaction
Central Nervous System
reverse transcriptase polymerase chain reaction
clones
Polymerase Chain Reaction
Messenger RNA
protein
Clone Cells

Keywords

  • C/EBP
  • CNS
  • Lymnaea
  • mRNA
  • qRT-PCR

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)

Cite this

Real-time quantitative RT-PCR method for estimation of mRNA level of CCAAT/enhancer binding protein in the central nervous system of Lymnaea stagnalis. / Hatakeyama, D.; Sadamoto, Hisayo; Ito, Etsuro.

In: Acta Biologica Hungarica, Vol. 55, No. 1-4, 2004, p. 157-161.

Research output: Contribution to journalArticle

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