Name: purification of mitochondria from yeast cells
Uploaded: 2009-08-24T16:00:00
Description: Watch this Scientific Journal Video about Purification of Mitochondria from Yeast Cells at JoVE.com

这项工作得到了CIHR和加拿大NSERC资助。 VIT是一个新的CIHR调查和Concordia大学基因组学研究主席，细胞生物学和老化。

材料和方法酵母菌株和生长条件。 丰富YEPD培养基中生长的野生型菌株BY4742（MATαhis3Δ1leu2Δ0lys2Δ0ura3Δ0）（1％酵母提取物，2％蛋白胨，2％葡萄糖） 。细胞培养在30 ° C旋转摇晃锥形瓶在200 RPM“瓶体积/中等音量”5:1的比例。 原油线粒体分数隔离<ol><li>长为48小时1升的野生型菌株BY4742文化</li><li>倒入500毫升NALGENE离心管文化。平衡离...

<ol>
	<li>Voet, D., Voet, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biochemistry.">Biochemistry.</a> , (2004).</li><li>Suen, D. -. F., Norris, K. L., Youle, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondrial+dynamics+and+apoptosis.">Mitochondrial dynamics and apoptosis.</a> Genes Dev. 22, 1577-1590 (2008).</li><li>Cheng, W. C., Berman, S. B., Ivanovska, I., Jonas, E. A., Lee, S. J., Chen, Y., Kaczmarek, L. K., Pineda, F. &. a. m. p. ;. a. m. p., Hardwick, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondrial+factors+with+dual+roles+in+death+and+survival.">Mitochondrial factors with dual roles in death and survival.</a> Oncogene. 25, 4697-4705 (2006).</li><li>Detmer, S. A., Chan, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functions+and+dysfunctions+of+mitochondrial+dynamics.">Functions and dysfunctions of mitochondrial dynamics.</a> Nat. Rev. Mol. Cell Biol. 8, 870-879 (2007).</li><li>McBride, H. M., Neuspiel, M., Wasiak, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondria:+more+than+just+a+powerhouse.">Mitochondria: more than just a powerhouse.</a> Curr. Biol. 16, R551-R560 (2006).</li><li>Balaban, R. S., Nemoto, S., Finkel, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondria,+oxidants,+and+aging.">Mitochondria, oxidants, and aging.</a> Cell. 120, 483-495 (2005).</li></ol>

这种方法使高产的隔离从酵母细胞中的纯线粒体。通过这种方法纯化的线粒体其他细胞器的污染基本上是免费的，其净化后保留其结构和功能的完整性。所述的方法产生的线粒体适用于无细胞重组基本线粒体过程。这些高度纯净的线粒体也可以用于线粒体结构和功能，线粒体蛋白质组和lipidome，和线粒体DNA的分析。

purification of mitochondria from yeast cells

线粒体是ATP生产的主要场所，在真核细胞非光合细胞的有氧代谢<sup&gt; 1</sup&gt;。这些复杂的细胞器在细胞凋亡中发挥重要作用<sup&gt; 2</sup&gt;，细胞的生存<sup&gt; 3</sup&gt;，哺乳动物的发展<sup&gt; 4</sup&gt;，神经细胞的发育和功能<sup&gt; 4</sup&gt;，细胞内的信号<sup&gt; 5</sup&gt;和长寿调控<sup&gt; 6</sup&gt;。我们的线粒体控制这些复杂的生物过程的理解，依靠强大的方法，以评估它们的形态，其蛋白质和脂质成分，它们的DNA的完整性，他们的许多重要职能。芽殖酵母<em&gt;酿酒酵母</em&gt;，一种基因和生物化学来操作带注释的基因组和蛋白质组定义良好的，单细胞真核生物是一个有价值的模型研究线粒体的重要生物功能的分子和细胞的内在机制，。对于这些类型的研究，这是至关重要的高纯度线粒体。在这里，我们提出了一个迅速和有效的方法纯化的酵母线粒体的详细描述。这种方法使其他细胞器的污染基本上是免费的，保留其净化后的结构和功能完整性的高度纯净的线粒体的隔离。通过这种方法纯化线粒体是适合无细胞重组基本线粒体过程，可用于线粒体结构和功能，线粒体的蛋白质和lipidome，和线粒体DNA的分析。

Watch this Scientific Journal Video about Purification of Mitochondria from Yeast Cells at JoVE.com

Purification of Mitochondria from Yeast Cells

我们描述了一个快速和有效的方法纯化的酵母线粒体<em&gt;酿酒酵母</em&gt;。这种方法使高产纯线粒体隔离，其他细胞器的污染基本上是免费的，并保留其净化后的结构和功能完整性。

从酵母细胞线粒体的纯化

Mitochondria are the main site of ATP production during aerobic metabolism in eukaryotic non-photosynthetic cells1. These complex organelles also play ...

Research

JoVE Journal

Biology

Isolation of CD4+ T cells from Mouse Lymph Nodes Using Miltenyi MACS Purification

分离的CD4 +从小鼠淋巴节点使用美天旎MACS纯化的T细胞

Isolation of cells from the primary source is a necessary step in many more complex protocols. Miltenyi offers kits to isolate cells from several ...

科研

生物学

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

无有机溶剂和醋酸，蛋白质与考马斯亮蓝G - 250凝胶染色

In classical protein staining protocols using Coomassie Brilliant Blue (CBB), solutions with high contents of toxic and flammable organic solvents ...

Purification of Progenitors from Skeletal Muscle

纯化的骨骼肌的先觉者

Skeletal muscle contains multiple progenitor populations of distinct embryonic origins and developmental potential. Myogenic progenitors, usually residing ...

Chromatographic Purification of Highly Active Yeast Ribosomes

高活性酵母核糖体的色谱分离纯化

Eukaryotic ribosomes are much more labile as compared to their eubacterial and archael counterparts, thus posing a significant challenge to researchers.  ...

Confocal Imaging of Single Mitochondrial Superoxide Flashes in Intact Heart or In Vivo

Confocal Imaging of Single Mitochondrial Superoxide Flashes in Intact Heart or In Vivo

单线粒体超氧闪烁在完整心脏或共聚焦成像<em&gt;在体内</em

Mitochondrion is a critical intracellular organelle responsible for energy production and intracellular signaling in eukaryotic systems. Mitochondrial ...

Isolation of mRNAs Associated with Yeast Mitochondria to Study Mechanisms of Localized Translation

Most of mitochondrial proteins are encoded in the nucleus and need to be imported into the organelle. Import may occur while the protein is synthesized ...

Rapid Isolation And Purification Of Mitochondria For Transplantation By Tissue Dissociation And Differential Filtration

Previously described mitochondrial isolation methods using differential centrifugation and/or Ficoll gradient centrifugation require 60 to 100 min to ...

Isolation and Functional Analysis of Mitochondria from Cultured Cells and Mouse Tissue

隔离和线粒体从培养细胞和小鼠组织功能分析

Comparison between two or more distinct groups, such as healthy vs. disease, is necessary to determine cellular status. Mitochondria are at the nexus of ...

Genome-wide Purification of Extrachromosomal Circular DNA from Eukaryotic Cells

从真核细胞染色体外的环状DNA基因组范围内的净化

Extrachromosomal circular DNAs (eccDNAs) are common genetic elements in Saccharomyces cerevisiae and are reported in other eukaryotes as well. EccDNAs ...

MitoCeption: Transferring Isolated Human MSC Mitochondria to Glioblastoma Stem Cells

MitoCeption：转让人离MSC线粒体脑胶质瘤干细胞

Mitochondria play a central role for cell metabolism, energy production and control of apoptosis. Inadequate mitochondrial function has been found ...