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

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.