A single-cell bioluminescence imaging system for monitoring cellular gene expression in a plant body.
A single-cell bioluminescence imaging system for monitoring cellular gene expression in a plant body.
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用于监测植物体内细胞基因表达的单细胞生物发光成像系统。
DOI：
10.1093/pcp/pct131
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Oyama T
中科院分区：
文献类型：
--
作者：
Muranaka T;Kubota S;Oyama T
基因表达是一个基本的细胞过程，表达动力学是生命科学的重要研究内容。我们成功地监测细胞基因表达的浮萍植物，浮萍，使用生物发光报告。使用粒子轰击，表皮和叶肉细胞用组成型[花椰菜花叶病毒35 S（CaMV 35 S）]和节律性[拟南芥CIRCADIAN CLOCK ASSOCIATED 1（AtCCA 1）]启动子控制下的荧光素酶基因（luc+）转染。使用EM-CCD（电子多电荷耦合器件）相机捕获生物发光图像。在单细胞水平上定量测量植物体内转染细胞的发光斑点。荧光强度变化超过1,000倍的范围内CaMV 35 S：：luc+转染细胞在同一植物体，并显示出对数正态样的频率分布。我们通过每小时捕获生物发光图像来监测光暗条件下的细胞基因表达。在每个监测程序中，成功地获得了叶中≥50个单个细胞的发光痕迹。在转染AtCCA 1：：luc+和CaMV 35 S：：luc+的细胞中分别观察到节律性和组成性发光行为。在每个AtCCA 1：：luc+引入的细胞中观察到昼夜节律，并具有可追踪的发光，并且在其节律波形中检测到轻微差异。因此，单细胞生物发光监测系统可用于表征植物体内细胞基因表达。
Gene expression is a fundamental cellular process and expression dynamics are of great interest in life science. We succeeded in monitoring cellular gene expression in a duckweed plant,Lemna gibba, using bioluminescent reporters. Using particle bombardment, epidermal and mesophyll cells were transfected with the luciferase gene (luc+) under the control of a constitutive [Cauliflower mosaic virus 35S(CaMV35S)] and a rhythmic [Arabidopsis thaliana CIRCADIAN CLOCK ASSOCIATED 1(AtCCA1)] promoter. Bioluminescence images were captured using an EM-CCD (electron multiply charged couple device) camera. Luminescent spots of the transfected cells in the plant body were quantitatively measured at the single-cell level. Luminescence intensities varied over a 1,000-fold range amongCaMV35S::luc+-transfected cells in the same plant body and showed a log-normal-like frequency distribution. We monitored cellular gene expression under light–dark conditions by capturing bioluminescence images every hour. Luminescence traces of ≥50 individual cells in a frond were successfully obtained in each monitoring procedure. Rhythmic and constitutive luminescence behaviors were observed in cells transfected withAtCCA1::luc+andCaMV35S::luc+, respectively. Diurnal rhythms were observed in everyAtCCA1::luc+-introduced cell with traceable luminescence, and slight differences were detected in their rhythmic waveforms. Thus the single-cell bioluminescence monitoring system was useful for the characterization of cellular gene expression in a plant body.
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DOI：
10.1046/j.1365-313x.2001.01063.x
发表时间：
2001-06
期刊：
The Plant journal : for cell and molecular biology
影响因子：
--
作者：
N. Sugiyama;T. Izawa;T. Oikawa;K. Shimamoto
通讯作者：
N. Sugiyama;T. Izawa;T. Oikawa;K. Shimamoto
影响因子：
1.9
作者：
Les, DH;Landolt, E;Crawford, DJ
通讯作者：
Crawford, DJ
影响因子：
4.9
作者：
T. Kondo;T. Tsudzuki
通讯作者：
T. Tsudzuki
浮萍 G3 衰老与气孔孔径之间缺乏相关性
影响因子：
4.9
作者：
Kwan Hee Park;K. Hahn;B. Kang
通讯作者：
B. Kang
DOI：
10.1111/j.1365-313x.2011.04707.x
发表时间：
2011-11
期刊：
The Plant journal : for cell and molecular biology
影响因子：
--
作者：
Esther Yakir;M. Hassidim;N. Melamed‐Book;Dror Hilman;Ido Kron;R. Green
通讯作者：
Esther Yakir;M. Hassidim;N. Melamed‐Book;Dror Hilman;Ido Kron;R. Green