Name: measuring replicative life span in the budding yeast
Uploaded: 2009-06-25T00:00:00
Description: Watch this Scientific Journal Video about Measuring Replicative Life Span in the Budding Yeast at JoVE.com

この作品は、エリソン医学財団からMKとBKKへの助成金によって支えられている。 MKは、高齢化のエリソン医学財団新奨学生です。私たちは、撮影時の支援をSoumya Kotireddyに感謝します。

パート1：複製の寿命解析のための準備菌株とプレートこのセクションでは、複製寿命の実験と寿命解析のための酵母細胞の調製のための固体YEPDプレートの準備について説明します。 <ol><li>適切な無菌操作を使用して、培養の酵母細胞および複製寿命解析のために使用されるYEPD寒天プレート（1％酵母エキス、2％バクト - ペプトン、2％寒天、2％グルコース）を準備。あなたは、寿命の...

<ol>
	<li>Kaeberlein, M., Burtner, C. R., Kennedy, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+developments+in+yeast+aging.">Recent developments in yeast aging.</a> PLoS Genet. 3, (2007).</li><li>Kaeberlein, M., Conn, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of models for human aging. , 109-120 (2006).</li><li>Smith, E. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+evidence+for+conserved+longevity+pathways+between+divergent+eukaryotic+species.">Quantitative evidence for conserved longevity pathways between divergent eukaryotic species.</a> Genome Res. 18, 564-570 (2008).</li><li>Steinkraus, K. A., Kaeberlein, M., Kennedy, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Replicative+aging+in+yeast:+the+means+to+the+end.">Replicative aging in yeast: the means to the end.</a> Annu Rev Cell Dev Biol. 24, 29-54 (2008).</li><li>Murakami, C. J., Burtner, C. R., Kennedy, B. K., Kaeberlein, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+for+high-throughput+quantitative+analysis+of+yeast+chronological+life+span.">A method for high-throughput quantitative analysis of yeast chronological life span.</a> J Gerontol A Biol Sci Med Sci. 63, 113-121 (2008).</li><li>Jiang, J. C., Jaruga, E., Repnevskaya, M. V., Jazwinski, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+intervention+resembling+caloric+restriction+prolongs+life+span+and+retards+aging+in+yeast.">An intervention resembling caloric restriction prolongs life span and retards aging in yeast.</a> Faseb J. 14, 2135-2137 (2000).</li><li>Kaeberlein, M., Kirkland, K. T., Fields, S., Kennedy, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sir2-independent+life+span+extension+by+calorie+restriction+in+yeast.">Sir2-independent life span extension by calorie restriction in yeast.</a> PLoS Biol. 2, E296-E296 (2004).</li><li>Lin, S. J., Defossez, P. A., Guarente, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Requirement+of+NAD+and+SIR2+for+life-span+extension+by+calorie+restriction+in+Saccharomyces+cerevisiae.">Requirement of NAD and SIR2 for life-span extension by calorie restriction in Saccharomyces cerevisiae.</a> Science. 289, 2126-2128 (2000).</li><li>Kaeberlein, M., Kennedy, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large-scale+identification+in+yeast+of+conserved+ageing+genes.">Large-scale identification in yeast of conserved ageing genes.</a> Mech Ageing Dev. 126, 17-21 (2005).</li><li>Kaeberlein, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+yeast+replicative+life+span+by+TOR+and+Sch9+in+response+to+nutrients.">Regulation of yeast replicative life span by TOR and Sch9 in response to nutrients.</a> Science. 310, 1193-1196 (2005).</li></ol>

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<td>Agar</td>
<td>Reagent</td>
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<td>Bacto-Peptone</td>
<td>Reagent</td>
<td>Fisher Scientific (BD Diagnostic Systems)</td>
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<td>&nbsp;</td>
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<td>Reagent</td>
<td>Fisher Scientific (BD Diagnostic Systems)</td>
<td>DF0886-17-0 (288620)</td>
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<td>Glucose</td>
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measuring replicative life span in the budding yeast

老化は細胞成分と死亡率をもたらす細胞内小器官の進行性の悪化によって特徴付け変性過程です。出芽酵母<em&gt;サッカロマイセスセレビシエ</em&gt;老化の生物学を研究するために広く使用されており、酵母の寿命のいくつかの決定は、ワーム、ハエ、マウスなどを含む多細胞真核生物で保存されることが示されている<sup&gt; 1</sup&gt;。容易に定量加齢に伴う表現型の欠如のために、酵母の老化は、一般的な用法の異なる2つの寿命のパラダイムで、さまざまなコンテキストで細胞の寿命を測定することによって、ほぼ独占的に検定されています<sup&gt; 2</sup&gt;。時系列寿命は、母細胞は非分裂、静止のような状態で生存できる時間の長さを指し、そして多細胞真核生物で有糸分裂後の細胞の老化のモデルとして機能するように提案されている。複製寿命は、対照的に、老化への事前の母細胞により産生される娘細胞の数を意味し、有糸分裂活性細胞の老化のモデルを提供すると考えられている。ここでは、出芽酵母の母細胞の複製寿命を測定するための一般的なプロトコルを提示する。複製寿命アッセイの目的は、何回、各母細胞の芽を決定することです。母と娘細胞は容易にそのようなツァイスのAxioscope 40または他の同等のモデルとして標準的な光学顕微鏡を（総合倍率160X）、使用経験のある研究者によって区別す​​ることができます。母細胞から娘細胞の物理的な分離は、光ファイバの針を装備した手動マイクロマニピュレーターを用いて実現されます。典型的な実験室酵母株は、母20〜30の娘細胞を生成し、一つの命のスパン実験では2-3週間必要です。

Watch this Scientific Journal Video about Measuring Replicative Life Span in the Budding Yeast at JoVE.com

Measuring Replicative Life Span in the Budding Yeast

この記事では、酵母の母細胞の複製寿命を測定するための一般的なプロトコルを提示する。

出芽酵母の複製寿命の測定

Aging is a degenerative process characterized by a progressive deterioration of cellular components and organelles resulting in mortality. The budding ...

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