Name: investigating tissue- and organ-specific phytochrome responses using facs-assisted cell-type specific expression profiling in arabidopsis thaliana
Uploaded: 2010-05-29T14:00:00
Description: Watch this Scientific Journal Video about Investigating Tissue- and Organ-specific Phytochrome Responses using FACS-assisted Cell-type Specific Expression Profiling in Arabidopsis thaliana at JoVE.com

蒙哥马利实验室工作，在植物中的光敏反应是支持由美国国家科学基金会（赠款。MCB - 0919100，以博莱霉素）和化学科学，地球科学和生物科学部，基础能源科学办公室，科学办公室，美国能源部能源（授予没有。DE FG02 91ER20021以博莱霉素）。我们感谢技术援助梅利莎惠特克在拍摄过程中，批判地阅读手稿，实验的援助，协助发展中国家和优化用共聚焦荧光激活细胞分选的援助拟南芥原生质体排序博士和梅林达框架协议的路易王博士斯蒂芬妮Costigan显微镜。我们感谢马琳卡梅伦图形设计援助和Karen鸟社论援助。

1。植物生长<ol><li>证实UAS -超视距X GAL4 - GFP增强陷阱线隔离4（对于总结见图1）和野生型或亲土壤播种，即〜2000年，每行消毒的种子。 </li><li>植物生长5周100μmolm-2 s - 1时的白色照明下对土壤在22℃，湿度70％ 。 </li></ol> 2。叶原生质体分离（改编自9 Denecke和Vitale的） <ol><li>为了分离原生质体，准备TEX的缓冲区。对于1公升TEX的缓冲区，掂量出了以下组件?...

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（1）通过微阵列基因表达分析表明，超过30％的基因在拟南芥幼苗中轻监管 11（2）已经确定了浩瀚的基因组编码的光信号转导有关的蛋白质的光敏色素信号级联12 ，13 。这样的实验表明，光诱导基因表达的快速和长期的变化。每个池的光敏色素，可控制的发展和适应性反应的一个子集。此外，它可能是下游的信号元件，在细胞和组织特异性的方式2，14，15中的光敏色素互...

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investigating tissue- and organ-specific phytochrome responses using facs-assisted cell-type specific expression profiling in arabidopsis thaliana

阵列的光介导植物的整个生命周期的发展和适应过程。植物利用光吸收分子，称为光感受器感知和适应光线。红/远红光的光吸收光敏感光器已被广泛的研究。光敏色素存在，在所有高等植物系统的独特和重叠的职能中，他们已经研究了蛋白质与家庭<sup&gt; 1</sup&gt;。光敏色素介导的光反应，从种子发芽到开花和衰老的范围，往往是本地化特定的植物组织或器官<sup&gt; 2</sup&gt;。尽管通过突变分析发现和澄清个人和冗余的光敏功能，photoperception和本地化池的光敏色素的分子机制不同地点的结论性报告，调解空间特定的光敏反应是有限的。我们设计实验的基础上的假设，调节组织器官特异性和光形态方面，而且本地化的光敏池从事细胞 - 细胞信号转导下游靶基因的不同的子集的特定网站的光敏photoperception。我们开发了一种生化方法选择性地减少在器官或组织特异性的方式在转基因植物的功能光敏。我们的研究是基于二部的增强子陷阱的方法，在上游激活序列（UAS）的元素的控制之下的一个基因的表达，反式转录激活GAL4<sup&gt; 3</sup&gt;。胆绿素还原酶（<em&gt;超视距</em&gt;默默地保持在GAL4式激活UAS -超视距母公司的情况下）根据无人机的控制基因<sup&gt; 4</sup&gt;。一个UAS超视距转线和一个GAL4 - GFP增强陷阱的具体表达结果之间的遗传杂交<em&gt;超视距</em&gt;在标记细胞的基因<em&gt;绿色荧光蛋白</em&gt;表达式<sup&gt; 4</sup&gt;。超视距积累在拟南芥植物光敏色素生色团缺乏的结果<em&gt;在植物</em<sup&gt; 5-7</sup&gt;。因此，我们制作了展品GAL4激活依赖的转基因植物<em&gt;超视距</em&gt;基因，导致光敏生化灭活，以及GAL4依赖<em&gt;绿色荧光蛋白</em&gt;表达式。光生物和分子遗传学分析<em&gt;超视距</em&gt;转基因株系产生洞察到组织和器官特异性的光敏色素介导，已与相应的网站photoperception相关的的反应<sup&gt; 4，7，8</sup&gt;。荧光激活细胞分选（FACS）GFP阳性，增强子陷阱诱导<em&gt;超视距</em&gt;表达植物原生质体细胞类型特异性基因通过表达谱芯片分析加上正在使用，以确定参与调解空间特定的光敏反应的假定下游靶基因。这项研究是扩大我们的网站光感的理解，通过光调控植物生长和发育的各种组织或器官的合作，推进复杂的光敏色素介导的分子清扫细胞 - 细胞信号传导机制。

Watch this Scientific Journal Video about Investigating Tissue- and Organ-specific Phytochrome Responses using FACS-assisted Cell-type Specific Expression Profiling in Arabidopsis thaliana at JoVE.com

Investigating Tissue- and Organ-specific Phytochrome Responses using FACS-assisted Cell-type Specific Expression Profiling in Arabidopsis thaliana

空间特定的光敏反应的分子基础正在调查中使用，表现出组织和器官特异性的光敏色素的不足的转基因植株。参展隔离特定的细胞芯片分析排序，是用来识别在特定空间的光敏反应有关的基因的荧光激活细胞诱导的光敏色素生色枯竭。

调查组织和器官特定的光敏色素反应，使用流式细胞仪辅助细胞类型特定的表达谱拟南芥</em

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Investigating Tissue- and Organ-specific Phytochrome Responses using FACS-assisted Cell-type Specific Expression Profiling in Arabidopsis thaliana

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