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

Tetsuya Miyamoto; Takahiro Nakamura; Issei Nagao; Junichi Obokata

doi:10.1007/BF02824017

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

Tetsuya Miyamoto, Takahiro Nakamura, Issei Nagao, Junichi Obokata

Research output: Contribution to journal › Article › peer-review

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 language	English
Pages (from-to)	101-107
Number of pages	7
Journal	Plant Molecular Biology Reporter
Volume	18
Issue number	2
DOIs	https://doi.org/10.1007/BF02824017
Publication status	Published - 2000
Externally published	Yes

All Science Journal Classification (ASJC) codes

Molecular Biology
Plant Science

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10.1007/BF02824017

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title = "Quantitative Analysis of Transiently Expressed mRNA in Particle-Bombarded Tobacco Seedlings",

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.",

author = "Tetsuya Miyamoto and Takahiro Nakamura and Issei Nagao and Junichi Obokata",

note = "Funding Information: was supported in part by the Grants-in-Aid from the Ministry of Education, Science and Culture of Japan.",

year = "2000",

doi = "10.1007/BF02824017",

language = "English",

volume = "18",

pages = "101--107",

journal = "Plant Molecular Biology Reporter",

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publisher = "Springer New York",

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TY - JOUR

T1 - Quantitative Analysis of Transiently Expressed mRNA in Particle-Bombarded Tobacco Seedlings

AU - Miyamoto, Tetsuya

AU - Nakamura, Takahiro

AU - Nagao, Issei

AU - Obokata, Junichi

N1 - Funding Information: was supported in part by the Grants-in-Aid from the Ministry of Education, Science and Culture of Japan.

PY - 2000

Y1 - 2000

N2 - 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.

AB - 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.

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DO - 10.1007/BF02824017

M3 - Article

AN - SCOPUS:0034561932

SN - 0735-9640

VL - 18

SP - 101

EP - 107

JO - Plant Molecular Biology Reporter

JF - Plant Molecular Biology Reporter

IS - 2

ER -

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

Abstract

All Science Journal Classification (ASJC) codes

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