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

Fingerprint

Glucuronidase
Seedlings
gene transfer
Tobacco
quantitative analysis
tobacco
reverse transcriptase polymerase chain reaction
Messenger RNA
seedlings
Plant Cells
introduced plants
RNA
cells
gene expression
Genes
Real-Time Polymerase Chain Reaction
proteins
Gene Expression
Polymerase Chain Reaction
methodology

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Plant Science

Cite this

Quantitative Analysis of Transiently Expressed mRNA in Particle-Bombarded Tobacco Seedlings. / Miyamoto, Tetsuya; Nakamura, Takahiro; Nagao, Issei; Obokata, Junichi.

In: Plant Molecular Biology Reporter, Vol. 18, No. 2, 01.01.2000, p. 101-107.

Research output: Contribution to journalArticle

Miyamoto, Tetsuya ; Nakamura, Takahiro ; Nagao, Issei ; Obokata, Junichi. / Quantitative Analysis of Transiently Expressed mRNA in Particle-Bombarded Tobacco Seedlings. In: Plant Molecular Biology Reporter. 2000 ; Vol. 18, No. 2. pp. 101-107.
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