Name: analyzing gene expression from marine microbial communities using environmental transcriptomics
Uploaded: 2009-02-18T00:00:00
Description: Watch this Scientific Journal Video about Analyzing Gene Expression from Marine Microbial Communities using Environmental Transcriptomics at JoVE.com

经费是由戈登和贝蒂摩尔基金会赠款和美国国家科学基金会的资助MCB - 0702125。

工作与RNA 由于RNase的是无处不在的mRNA迅速降解，在核糖核酸酶自由的环境中工作，必须遵循和样品应尽快以下尽可能收集加工或腌制的标准预防措施。 第1部分：环境RNA集合 （旨在收集在0.2 - 3.0微米大小的一小部分生物量） 用品需要： 的Masterflex管 蠕动泵 3微米高容量褶囊式过滤器 苏泊尔0.22微米或聚碳?...

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	<li>Liang, P., Pardee, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Science. 257 (5072), 967-967 (1992).</li><li>Belasco, J. G., Belasco, J. G., Brawerman, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+of+messenger+RNA+stability.">Control of messenger RNA stability.</a> , 3-11 (1993).</li><li>Poretsky, R. S., Bano, N., Buchan, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> 71 (7), 4121-4121 (2005).</li><li>Gelder, R. N. V., von Zastrow, M. E., Yool, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Natl. Acad. Sci. USA. 87 (5), 1663-1663 (1990).</li><li>Margulies, M., Egholm, M., Altman, W. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Nature. 437 (7057), 376-376 (2005).</li><li>Frias-Lopez, J., Shi, Y., Tyson, G. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Natl. Acad. Sci. USA. 105 (10), 3805-3805 (2008).</li><li>Poretsky, R. S., Hewson, I., Sun, S., Allen, A. E., Zehr, J. P., Moran, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Environ Microbiol. 11 (6), 1358-1375 (2009).</li></ol>

基因表达的天然微生物群落的调查，近年来已成为共同探索微生物的生态作用和功能的一种手段。海洋微生物的检测，量化，并表征结合使用，可用于基因表达分析链接系统发育功能的天然微生物群落。为目标的功能基因表达的许多技术，依赖于特异性探针或基因的已知序列设计的引物。相比之下，环保转录可用于研究基因的表达没有施加的限制，现有的序列数据，并正在积极表达这些基因的偏?...

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<td>mRNA-ONLY Prokaryotic mRNA Isolation Kit</td>
<td>kit</td>
<td>Epicentre</td>
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<td>MICROBExpress Bacterial mRNA Enrichment Kit</td>
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<td>Universal RiboClone cDNA Synthesis System</td>
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<td>Wizard DNA Clean-Up System</td>
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analyzing gene expression from marine microbial communities using environmental transcriptomics

类似于宏基因组学，环境转录（metatranscriptomics）检索和环境微生物组合没有先验知识的社会可能是哪些基因表达序列的mRNA。因此，它提供了最不偏不倚的角度，对社区的基因表达<em&gt;原位</em&gt;。环境转录协议在技术上是困难的，因为原核生物mRNA的普遍缺乏的poly（A）尾巴，使真核细胞的消息隔离相对简单<sup&gt; 1</sup&gt;因为相对较短的mRNA半衰期<sup&gt; 2</sup&gt;。此外，mRNA的是远远低于在总RNA提取的rRNAs丰富，因此rRNA的背景往往淹没mRNA的信号。但是，为克服这些困难，一些技术最近被开发出来。最近描述一个用于分析环境创建克隆库，采用随机引物进行反向转录和健全环境的mRNA转录的过程是成功的在两个不同的自然环境，但结果被用于启动cDNA合成的随机引物的选择偏颇<sup&gt; 3</sup&gt;。线性扩增的mRNA的进展，避免随机引物扩增步骤的需要，并作出有可能使用较少的起始原料，从而降低了样品的收集和处理时间，从而最大限度地减少RNA降解<sup&gt; 4</sup&gt;。<em在体外</em放大mRNA的转录方法涉及polyadenylating的mRNA，并纳入到3月末的成绩单T7启动子。扩增RNA（ARNA），然后可以被转换为双用随机六聚体的双链cDNA，并直接测序由焦磷酸测序<sup&gt; 5</sup&gt;。第一次使用这种方法的一个站ALOHA表明微生物群落的基因表达特征的效用<sup&gt; 6</sup&gt;。

Watch this Scientific Journal Video about Analyzing Gene Expression from Marine Microbial Communities using Environmental Transcriptomics at JoVE.com

Analyzing Gene Expression from Marine Microbial Communities using Environmental Transcriptomics

我们提出了一个方法生成环境的mRNA的cDNA。在一般情况下，总RNA是第一次从环境中收集，rRNA的选择性去除，mRNA的选择性扩增，并从丰富的mRNA池合成cDNA测序。使用标准的生物信息学技术确定基因表达，恢复序列，可以注明。

社区环境转录从海洋微生物基因表达分析

Analogous to metagenomics, environmental transcriptomics (metatranscriptomics) retrieves and sequences environmental mRNAs from a microbial assemblage ...

Research

JoVE Journal

Biology

Microbial Communities in Nature and Laboratory - Interview

微生物群落，在自然和实验室 - 面试

科研

生物学

Biology of Microbial Communities  - Interview

微生物群落的生物 - 面试

Bacterial Gene Expression Analysis Using Microarrays

使用微阵列的细菌基因表达分析

The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation

主电穿孔的小鼠骨髓造血细胞培养制备

It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser ...

Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency

使用基因脉冲MXcell电穿孔系统主细胞的转染效率高

Using an Automated Cell Counter to Simplify Gene Expression Studies: siRNA Knockdown of IL-4 Dependent Gene Expression in Namalwa Cells

使用自动细胞计数器简化基因表达研究：在Namalwa细胞的IL - 4依赖的基因表达的siRNA击倒

The use of siRNA mediated gene knockdown is continuing to be an important tool in studies of gene expression.  siRNA studies are being conducted not only ...

In vitro Reconstitution of the Active T. castaneum Telomerase

In vitro Reconstitution of the Active T. castaneum Telomerase

在体外重组活跃T。谷盗端粒酶

Efforts to isolate the catalytic subunit of telomerase, TERT, in sufficient quantities for structural studies, have been met with limited success for more ...

Extraction of High Molecular Weight DNA from Microbial Mats

超高分子量DNA提取微生物席

Successful and accurate analysis and interpretation of metagenomic data is dependent upon the efficient extraction  of high-quality, high molecular weight ...

Concentration of Metabolites from Low-density Planktonic Communities for Environmental Metabolomics using Nuclear Magnetic Resonance Spectroscopy

代谢物的浓度从低密度浮游社区的环境代谢，利用核磁共振光谱

Environmental metabolomics is an emerging field that is promoting new understanding in how organisms respond to and interact with the environment and each ...

Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides

作为一种工具来隔离核糖体的束缚多肽SECM逮捕序列

Extensive research has provided ample evidences suggesting that protein folding in the cell is a co-translational process1-5. However, the exact pathway ...