Quantitative Analysis of Transiently Expressed mRNA in Particle-Bombarded Tobacco Seedlings

Tetsuya Miyamoto, Takahiro Nakamura, Issei Nagao, Junichi Obokata

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

This study demonstrates that quantitative RT-PCR can be used to measure transient expression of genes introduced into plant cells by particle bombardment. An expression construct for β-glucuronidase was introduced into tobacco seedlings by particle bombardment followed by real-time quantitative RT-PCR of β-glucuronidase mRNA using a fluorogenic TaqMan probe. β-glucuronidase mRNA expression peaked within 2 h after gene transfer. β-glucuronidase protein activity was maximally elevated in plant cells 8 h after gene transfer and remained elevated for up to 50 h. This method is very sensitive, quantitating the target GUS transcript in 10 pg total RNA.

Original languageEnglish
Pages (from-to)101-107
Number of pages7
JournalPlant Molecular Biology Reporter
Volume18
Issue number2
DOIs
Publication statusPublished - Jan 1 2000
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Plant Science

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