Reporter gene expression at single-cell level characterized with real-time RT-PCR, chemiluminescence, fluorescence, and electrochemical imaging

被引：9

作者：

Shiku, Hitoshi ^{[1
]}

Okazaki, Daisuke ^{[1
]}

Suzuki, Junya ^{[1
]}

Takahashi, Yasufumi ^{[1
]}

Murata, Tatsuya ^{[1
]}

Akita, Hidetaka ^{[2
]}

Harashima, Hideyoshi ^{[2
]}

Ino, Kosuke ^{[1
]}

Matsue, Tomokazu ^{[1
]}

机构：

[1] Tohoku Univ, Grad Sch Environm Studies, Sendai, Miyagi 9808579, Japan

[2] Hokkaido Univ, Fac Pharmaceut Sci, Kita Ku, Sapporo, Hokkaido 0600812, Japan

来源：

FEBS LETTERS | 2010年 / 584卷 / 18期

关键词：

Transcription; Translation; Single-cell analysis; Reporter assay; EMBRYONIC STEM-CELLS; YEAST SCHIZOSACCHAROMYCES-POMBE; GLOBAL ANALYSIS; ADHERENT CELLS; MESSENGER-RNA; PROTEIN; LOCALIZATION; SPHEROIDS; PROCESSOR; SYSTEM;

D O I：

10.1016/j.febslet.2010.08.008

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

mRNA from single cells was quantified using real-time RT-PCR after recording the address and reporter protein activity with chemiluminescence, fluorescence, and electrochemical techniques, using luciferase, green fluorescent protein, and secreted alkaline phosphatase. mRNA copy number ranging from below 103 to 107 in single cells showed a lognormal distribution for both externally introduced reporter genes and internally expressed genes. The fluctuation in the gene expression decreased with the increase of the number of cells picked but did not decrease with the increase of mRNA copy number per cell. We found that the correlation coefficients for mRNA and protein expression in logarithmic plot at single-cell level were much lower than 1.00. (c) 2010 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

引用

页码：4000 / 4008

页数：9

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