Name: floral-dip transformation of arabidopsis thaliana to examine ptso2::&beta;-glucuronidase reporter gene expression
Uploaded: 2010-06-11T17:00:00
Description: Watch this Scientific Journal Video about Floral-dip Transformation of Arabidopsis thaliana to Examine pTSO2::ÃŽÂ²-glucuronidase Reporter Gene Expression at JoVE.com

我们感谢春新王建设pTSO2：：GUS和图1，德特勒夫威格尔的照片分享有益的意见的GUS万无一失的协议，帕哈Sijacic和考特尼荷伦德。 Z. L S实验室的研究是由美国国家科学基金会（IOB0616096和MCB0744752）的支持。 ZL是由美国马里兰大学农业试验站的部分支持。

一，花香浸拟南芥的转化<ol><li>改造前约1个月，大约有10-15％的盆栽植物拟南芥种子播撒到4至8盆（5平方英寸的花盆）。与正常生育的植物，4盆植物每质粒的构建就足够了。如果一个人改造拟南芥突变生育率下降，增加相应的盆数。 </li><li>植物生长标准条件下（16小时光照，8小时黑暗，在22 ° C）。如果较短的一天或更低的温度下生长的植物，他们将需要更长的时间，花和更...

<ol>
	<li>Clough, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Floral+dip:+agrobacterium-mediated+germ+line+transformation.">Floral dip: agrobacterium-mediated germ line transformation.</a> Methods Mol Biol. 286, 91-91 (2005).</li><li>Clough, S. J., Bent, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Floral+dip:+a+simplified+method+for+Agrobacterium-mediated+transformation+of+Arabidopsis+thaliana.">Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana.</a> Plant J. 16 (6), 735-735 (1998).</li><li>Wang, C., Liu, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arabidopsis+ribonucleotide+reductases+are+critical+for+cell+cycle+progression,+DNA+damage+repair,+and+plant+development.">Arabidopsis ribonucleotide reductases are critical for cell cycle progression, DNA damage repair, and plant development.</a> Plant Cell. 18 (2), 350-350 (2006).</li></ol>

转化效率是由许多不同的因素，下面讨论的： <ol><li>植物的健康和他们的年龄是最重要的。约一个月的老厂生产大量未成熟的花芽，是理想的。修剪主芽，以鼓励中学改造前3-4天拍摄形成会增加转化效率。老厂将产生转化，但在较低的利率。 </li><li>生育的植物是重要的。如果一个人改造与生育率下降的突变体，植物将需要更多的浸渍。 </li><li> 0.3％Silwet L - 77渗透缓冲区中的表面活性剂被认为?...

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<td>6-benzyladenine</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>852430</td>
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<td>Silwet-77</td>
<td>&nbsp;</td>
<td>Crompton Corp.</td>
<td>&nbsp;</td>
<td>Toxic, wear gloves</td>
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<td>Murashige and Skoog Basal medium (MS)</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>M5519</td>
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<td>Gamborg's vitamin solution 1000X</td>
<td>&nbsp;</td>
<td>Sigma</td>
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<td>X-Gluc (5-bromo-4-chloro-3-indolyl &beta;-D-glucuronide cyclohexamine salt</td>
<td>&nbsp;</td>
<td>Biosynth</td>
<td>B-7300</td>
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<td>&nbsp;</td>
<td>Fisher</td>
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floral-dip transformation of arabidopsis thaliana to examine ptso2::&beta;-glucuronidase reporter gene expression

一个有机体，引入外源基因的能力，是现代生物学和生物技术的基础。在该模型中的开花植物<em&gt;拟南芥</em花浸变换方法<sup&gt; 1-2</sup&gt;已取代所有以前的方法，因为它的简单，效率，成本低。具体来说，年轻开花的芽<em&gt;拟南芥</em&gt;植物的溶液浸泡<em&gt;农杆菌</em&gt;携带特定的质粒结构。浸渍后，植物恢复到正常的生长和产量的种子，很小的比例，其中与外源基因的转化，可以选择含抗生素的培养基上。这花香浸方法，大大地促进<em&gt;拟南芥</em&gt;研究大大促进了我国植物基因功能的理解。在这项研究中，我们使用花香浸法记者基因改造，<em&gt;β-葡萄糖醛酸酶（GUS）的</em&gt;下，控制<em&gt; TSO2</em&gt;推动者。<em&gt; TSO2</em&gt;，核苷酸还原酶（RNR）小亚基编码<sup&gt; 3</sup&gt;，是一个细胞周期调控基因在细胞周期S期dNDP合成的必要条件。审核<em&gt; GUS</em&gt;在转基因的表达<em&gt;拟南芥</em&gt;苗显示，<em&gt; TSO2</em&gt;表示积极分裂组织。报告的实验方法和材料可以很容易地适应不仅为研究，而且还对教育，在高中和大学水平。

Watch this Scientific Journal Video about Floral-dip Transformation of Arabidopsis thaliana to Examine pTSO2::ÃŽÂ²-glucuronidase Reporter Gene Expression at JoVE.com

Floral-dip Transformation of Arabidopsis thaliana to Examine pTSO2::ÃƒÅ½Ã‚Â²-glucuronidase Reporter Gene Expression

本文介绍了花卉浸方法<em&gt;农杆菌</em&gt;介导的转化<em&gt;拟南芥</em&gt;。通过引入一个细胞周期调节的发起人，记者，<em&gt; pTSO2：β-葡萄糖醛酸酶（GUS）的</em&gt;，到<em&gt;拟南芥</em&gt;，我们演示了如何检测到GUS基因在转基因苗记者表达。

花香浸改造拟南芥检查 pTSO2：β-葡萄糖醛酸酶报告基因的表达

The ability to introduce foreign genes into an organism is the foundation for modern biology and biotechnology. In the model flowering plant Arabidopsis ...

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Floral-dip Transformation of Arabidopsis thaliana to Examine pTSO2::&beta;-glucuronidase Reporter Gene Expression

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