Name: assessment of de novo protein synthesis rates in caenorhabditis elegans
Uploaded: 2020-09-12T13:00:00
Description: Watch this Scientific Journal Video about Assessment of de novo Protein Synthesis Rates in Caenorhabditis elegans at JoVE.com

我们感谢查尼奥塔基斯 M. 和库纳基斯 K. 的录像录制和编辑。K.P. 由希腊研究和创新基金会（HFRI）和研究与技术总秘书处（GSRT）提供赠款。N.T. 的资金来自欧洲研究理事会 （ERC – GA695190 – MANNA）、欧盟委员会框架方案和希腊教育部的赠款。

注：对荧光团的转录和转化融合都可用于评估多个组织中的诺沃蛋白转化。蛋白质合成率可以评估多个蛋白质家族，本地化在不同的细胞隔间，包括细胞质，线粒体和核7。以下菌株用于监测全球和神经元蛋白合成率，N2;Ex[pife-2GFP， pRF4]， ife-2 （ok306）;Ex[pife-2GFP， pRF4]， N2;Ex[punc-119GFP， pRF4]， edc-3 （ok1427...

<ol>
	<li>Syntichaki, P., Troulinaki, K., Tavernarakis, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=eIF4E+function+in+somatic+cells+modulates+ageing+in+Caenorhabditis+elegans.">eIF4E function in somatic cells modulates ageing in Caenorhabditis elegans.</a> Nature. 445 (7130), 922-926 (2007).</li><li>Charmpilas, N., Daskalaki, I., Papandreou, M. E., Tavernarakis, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protein+synthesis+as+an+integral+quality+control+mechanism+during+ageing.">Protein synthesis as an integral quality control mechanism during ageing.</a> Ageing Research Reviews. 23, 75-89 (2015).</li><li>Li, G. W., Burkhardt, D., Gross, C., Weissman, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantifying+absolute+protein+synthesis+rates+reveals+principles+underlying+allocation+of+cellular+resources.">Quantifying absolute protein synthesis rates reveals principles underlying allocation of cellular resources.</a> Cell. 157 (3), 624-635 (2014).</li><li>Dermit, M., Dodel, M., Mardakheh, F. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+monitoring+and+measurement+of+protein+translation+in+time+and+space.">Methods for monitoring and measurement of protein translation in time and space.</a> Molecular Biosystems. 13 (12), 2477-2488 (2017).</li><li>Iwasaki, S., Ingolia, N. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Growing+Toolbox+for+Protein+Synthesis+Studies.">The Growing Toolbox for Protein Synthesis Studies.</a> Trends in Biochemical Sciences. 42 (8), 612-624 (2017).</li><li>Corsi, A. K., Wightman, B., Chalfie, 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+Transparent+Window+into+Biology:+A+Primer+on+Caenorhabditis+elegans.">A Transparent Window into Biology: A Primer on Caenorhabditis elegans.</a> Genetics. 200 (2), 387-407 (2015).</li><li>Kourtis, N., Tavernarakis, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell-specific+monitoring+of+protein+synthesis+in+vivo.">Cell-specific monitoring of protein synthesis in vivo.</a> PLoS One. 4 (2), 4547 (2009).</li><li>Parker, R., Sheth, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=P+bodies+and+the+control+of+mRNA+translation+and+degradation.">P bodies and the control of mRNA translation and degradation.</a> Molecular Cell. 25 (5), 635-646 (2007).</li><li>Rieckher, M., Markaki, M., Princz, A., Schumacher, B., Tavernarakis, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maintenance+of+Proteostasis+by+P+Body-Mediated+Regulation+of+eIF4E+Availability+during+Aging+in+Caenorhabditis+elegans.">Maintenance of Proteostasis by P Body-Mediated Regulation of eIF4E Availability during Aging in Caenorhabditis elegans.</a> Cell Reports. 25 (1), 199-211 (2018).</li><li>Reits, E. A., Neefjes, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+fixed+to+FRAP:+measuring+protein+mobility+and+activity+in+living+cells.">From fixed to FRAP: measuring protein mobility and activity in living cells.</a> Nature Cell Biology. 3 (6), 145-147 (2001).</li><li>Keith, S. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Graded+Proteasome+Dysfunction+in+Caenorhabditis+elegans+Activates+an+Adaptive+Response+Involving+the+Conserved+SKN-1+and+ELT-2+Transcription+Factors+and+the+Autophagy-Lysosome+Pathway.">Graded Proteasome Dysfunction in Caenorhabditis elegans Activates an Adaptive Response Involving the Conserved SKN-1 and ELT-2 Transcription Factors and the Autophagy-Lysosome Pathway.</a> PLoS Genetics. 12 (2), 1005823 (2016).</li><li>Kumsta, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+autophagy+receptor+p62/SQST-1+promotes+proteostasis+and+longevity+in+C.+elegans+by+inducing+autophagy.">The autophagy receptor p62/SQST-1 promotes proteostasis and longevity in C. elegans by inducing autophagy.</a> Nature Communications. 10 (1), 5648 (2019).</li><li>Gregersen, N., Bolund, L., Bross, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protein+misfolding,+aggregation,+and+degradation+in+disease.">Protein misfolding, aggregation, and degradation in disease.</a> Molecular Biotechnology. 31 (2), 141-150 (2005).</li></ol>

蛋白质合成调制对机体平衡至关重要。在衰老期间，全球和特定的蛋白质合成受到干扰。最近的研究表明，蛋白质转化平衡直接控制衰老和衰老不仅仅是衰老过程的副产品。特别是，真核启动因子4E（eIF4E）等翻译机制的核心成分，在翻译启动过程中促进mRNA封顶，从而降低依赖帽的蛋白质转化率，诱发氧化应激，加速衰老过程1。此外，神经元特异性EDC-3，它与mRNA处理体、P体相互...

蛋白质的合成和降解对于有机体平衡至关重要。许多与年龄相关的疾病是由有缺陷的蛋白质生产1，2,引起的。为了测量全球蛋白质的转化率，有生化技术，如核糖体分析，其中包括核糖体保护mRNA片段的深度测序，以监测表达以及新的蛋白质合成3。这种方法除了间接读出转化率外，因为增加RNA与核糖体的关联并不一定意味着翻译增加，因此具有技术缺点，例如成本高和大量起始材料的需求。另一方面，基于蛋白质组学的方法允许通过脉冲代谢分析直接蛋白质定量，然后进行质谱分析4，5。,5然而，这是一种半定量方法，具有有限的时间分辨率，在体内不容易使用。此外，蛋白质的标签可以在感兴趣的动物/组织中不均匀分布。重要的是，这两种方法可以分别隐藏整个动物或组织中蛋白质转化率的组织特异性或细胞特异性变异。
卡诺哈布迪蒂斯埃利 根是一种易于使用的模型生物体，可以大量生长6。此外，其遗传的适用性和透明度允许在体内进行活成像。该协议描述了光漂白（FRAP）后使用荧光回收检测蛋白质合成率的方法。我们利用荧光蛋白的转基因表达，无论是整个生物体还是特定的组织/细胞。转基因动物可以使用基因的启动子表达GP，该基因被广泛/全球表达，或者使用组织特异性启动子来靶向特定细胞类型。此技术可以扩展到特定启动子，以检查特定蛋白质的蛋白质合成率。

<table>
	<tbody>
		<tr>
			<td>Agar</td>
			<td>Sigma-Aldrich</td>
			<td>5040</td>
			<td></td>
		</tr>
		<tr>
			<td>Agarose</td>
			<td>Biozym</td>
			<td>8,40,004</td>
			<td></td>
		</tr>
		<tr>
			<td>Agarose pads 2%</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Calcium chloride dehydrate (CaCl2&#8729;2H2O)</td>
			<td>Sigma-Aldrich</td>
			<td>C5080</td>
			<td></td>
		</tr>
		<tr>
			<td>Cholesterol</td>
			<td>SERVA Electrophoresis</td>
			<td>17101.01</td>
			<td></td>
		</tr>
		<tr>
			<td>cycloheximide</td>
			<td>Sigma-Aldrich</td>
			<td>C-7698</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting stereomicroscope</td>
			<td>Nikon Corporation</td>
			<td></td>
			<td>SMZ645</td>
		</tr>
		<tr>
			<td>edc-3(ok1427);Ex[punc-119GFP, pRF4]</td>
			<td>Tavernarakis lab</td>
			<td></td>
			<td>Maintain animals at 20 &#176;C</td>
		</tr>
		<tr>
			<td>Epifluorescence microscope</td>
			<td>Zeiss</td>
			<td></td>
			<td>Axio Imager Z2</td>
		</tr>
		<tr>
			<td>Escherichia coli OP50 strain</td>
			<td>Caenorhabditis Genetics Center (CGC)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescence dissecting stereomicroscope</td>
			<td>Zeiss</td>
			<td></td>
			<td>SteREO Lumar V12</td>
		</tr>
		<tr>
			<td>Greiner Petri dishes (60 x 15 mm)</td>
			<td>Sigma-Aldrich</td>
			<td>P5237</td>
			<td></td>
		</tr>
		<tr>
			<td>ife-2(ok306);Ex[pife-2GFP, pRF4]</td>
			<td>Tavernarakis lab</td>
			<td></td>
			<td>Maintain animals at 20 &#176;C</td>
		</tr>
		<tr>
			<td>image analysis software</td>
			<td>Fiji</td>
			<td></td>
			<td>https://fiji.sc</td>
		</tr>
		<tr>
			<td>KH2PO4</td>
			<td>EMD Millipore</td>
			<td>1,37,010</td>
			<td></td>
		</tr>
		<tr>
			<td>K2HPO4</td>
			<td>EMD Millipore</td>
			<td>1,04,873</td>
			<td></td>
		</tr>
		<tr>
			<td>LB liquid medium</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>M9 buffer</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Magnesium sulfate (MgSO4)</td>
			<td>Sigma-Aldrich</td>
			<td>M7506</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope slides (75 x 25 x 1 mm)</td>
			<td>Marienfeld, Lauda-Koenigshofen</td>
			<td>10 006 12</td>
			<td></td>
		</tr>
		<tr>
			<td>Microsoft Office 2011 Excel software package</td>
			<td>Microsoft Corporation, Redmond, USA</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>N2;Ex[pife-2GFP; pRF4]</td>
			<td>Tavernarakis lab</td>
			<td></td>
			<td>Maintain animals at 20 &#176;C</td>
		</tr>
		<tr>
			<td>N2;Ex[psod-3GFP; pRF4]</td>
			<td>Tavernarakis lab</td>
			<td></td>
			<td>Maintain animals at 20 &#176;C</td>
		</tr>
		<tr>
			<td>N2;Ex[punc-119GFP; pRF4]</td>
			<td>Tavernarakis lab</td>
			<td></td>
			<td>Maintain animals at 20 &#176;C</td>
		</tr>
		<tr>
			<td>Na2HPO4</td>
			<td>EMD Millipore</td>
			<td>1,06,586</td>
			<td></td>
		</tr>
		<tr>
			<td>Nematode growth medium (NGM) agar plates</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Nystatin stock solution</td>
			<td>Sigma-Aldrich</td>
			<td>N3503</td>
			<td></td>
		</tr>
		<tr>
			<td>Peptone</td>
			<td>BD, Bacto</td>
			<td>211677</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate buffer</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium chloride (NaCl)</td>
			<td>EMD Millipore</td>
			<td>1,06,40,41,000</td>
			<td></td>
		</tr>
		<tr>
			<td>Standard equipment for preparing agar plates (autoclave, Petri dishes, etc.)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Standard equipment for maintaining worms (platinum wire pick, incubators, etc.).</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>statistical analysis software</td>
			<td>GraphPad Software Inc., San Diego, USA</td>
			<td></td>
			<td>GraphPad Prism software package</td>
		</tr>
		<tr>
			<td>Streptomycin</td>
			<td>Sigma-Aldrich</td>
			<td>S6501</td>
			<td></td>
		</tr>
		<tr>
			<td>UV Crosslinker</td>
			<td>Vilber Lourmat BIO-LINK BLX 254</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Worm pick</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

assessment of de novo protein synthesis rates in caenorhabditis elegans

保持健康的蛋白质组对细胞和有机体平衡至关重要。蛋白质转化控制和降解之间的平衡的扰动引发了许多与年龄相关的疾病。蛋白位虫质量控制机制的减少是老龄化的一个标志。检测新蛋白合成的生化方法仍然有限，有几个缺点，不能在活细胞或动物中进行。 卡诺哈布迪蒂斯，透明 和容易转基因，是一个极好的模型，以监测蛋白质合成率使用成像技术。在这里，我们介绍并描述了一种利用光漂白后荧光回收（FRAP）测量体内新蛋白合成的方法。在特定细胞或组织中表达荧光蛋白的转基因动物被强大的光源照射，从而产生荧光光漂白。反过来，荧光恢复的评估意味着新的蛋白质合成在细胞和/或组织感兴趣。因此，将转基因线虫、遗传和/或药理干预与蛋白质合成率的活成像相结合，可以揭示调解年龄依赖性蛋白酶崩溃的机制。

利用此处介绍的程序，使用野生型和突变转基因线虫表示以下体细胞学记者，pife-22GFP，psod-3-3GFP和 punc-119GFP，根据各自的方案评估蛋白质合成速率。特别是，野生型和 ife-2突变蠕虫使用ife-2促进器在其躯体组织中表达细胞质 GFP，在光漂白后和恢复后 5 小时立即进行比较。代表性图像和定量显示，野生类动物完...

Watch this Scientific Journal Video about Assessment of de novo Protein Synthesis Rates in Caenorhabditis elegans at JoVE.com

Assessment of de novo Protein Synthesis Rates in Caenorhabditis elegans

在这里，我们介绍和描述一种非放射性和非侵入性的方法，以评估体内的去新蛋白合成，利用线虫 卡诺 哈布迪炎和荧光漂白后恢复（FRAP）。该方法可与遗传和/或药理学屏幕相结合，识别蛋白质合成的新调制器。

卡诺哈布迪蒂斯 伊利根斯新蛋白合成率评估

Maintaining a healthy proteome is essential for cell and organismal homeostasis. Perturbation of the balance between protein translational control and ...

蛋白质合成的速度在疾病和衰老期间受到干扰。光漂白后荧光恢复（FRAP）允许测量体内蛋白质合成的速率。我们使用透明C.elegans蠕虫（表达GP）作为模型，监测光漂白后的新蛋白质合成。我们提供实用的指南，通过使用转基因蠕虫测量荧光恢复，转基因蠕虫在不同的启动子下表达GP，无论是广泛表达的还是特定的细胞或组织中。同时，该方法提供实时蛋白质合成速度监测。使用解剖立体显微镜评估野生型和突变转基因线虫的发育阶段和生长。挑选10个L4幼虫的野生型和突变转基因动物携带所需的荧光记者到线虫生长介质板与大肠杆菌种子。在20摄氏度的标准温度下孵育和生长线虫。四天后，该板块含有转基因蠕虫的混合种群。在新鲜种子OP50 NGM板上选择并转移每种菌株的15个L4幼虫。进行FRAP测定，并在成年第一天监测蛋白质合成率。第二天，准备并使用含有环氧胺的NGM板作为正对。通过暴露种子的NGM板15分钟与紫外线杀死细菌。在细菌种子板的顶部添加环氧胺，在总容量中每毫升最终浓度为 500 微克。让盘子干燥。转移在车辆和含环氧胺板上表达GP的转基因线虫。在20摄氏度的标准温度下孵育动物两小时。挑选并转移一天的成年转基因动物到个别NGM板种子与20微升OP50下降在中心。取下盖子，将每个板放在荧光显微镜的 20H 客观透镜下。在光漂白之前对参考图像进行聚焦和捕获。光漂白每个样品10分钟。光漂白后捕获图像。将动物放在单独的NGM盘子里，让它们恢复。在荧光立体显微镜下，每一小时对每个荧光测量仪进行一次图像和记录，至少6小时。准备2%的加糖垫，并在加糖垫的中心添加10微升的M9缓冲液。将表达泛神经元细胞质GP的5个转基因线虫转移到一滴M9缓冲液中。用睫毛传播液体。由于 M9 吸收剂进入阿加，动物在两分钟内表现出的运动减少。使用睫毛选择改变线虫的位置。将样品放在荧光显微镜的 40H 客观透镜上，无需使用盖滑。聚焦并捕获参考图像。光漂白目标感兴趣区域 90 秒。光漂白后捕获图像。在光漂白线虫上添加 10 微升 M9 缓冲液。让动物们恢复五分钟。使用睫毛拾取器或移液器将线虫转移到单个 NGM 板，在中心放置 20 微升 OP50 滴。在荧光立体显微镜下，每一小时拍摄一次每个样品的图像。使用 fe-2 启动子在细胞体组织中表达细胞质 GFP 的野生型和 fe-2 突变蠕虫在光漂白之前、光漂白后立即和恢复后 5 小时进行比较。野生类动物完全恢复，而fe-2突变体有迷你恢复能力。因此，野生型蠕虫在光漂白后启动正常蛋白质合成，而缺乏mRNA翻译启动因子fe-2的蠕虫则无法这样做，这表明荧光恢复率说明了体内蛋白质合成的速率。Cycloheximide是mRNA转化的特定抑制剂，可用作蛋白质转化抑制的阳性控制。事实上，在启动子下表达细胞质GP的环氧异酰胺处理转基因动物在光漂白时不会恢复其荧光。mRNA处理体影响蛋白质转化率。野生型和edc-3突变线虫表达GP泛神经元检查其恢复能力在目标光漂白在头部区域。与野生型蠕虫相比，edc-3缺陷动物的荧光恢复速度要慢得多。蛋白质合成调制对机体平衡至关重要。在衰老期间，全球和特定的蛋白质合成受到干扰。蛋白质转化平衡直接控制衰老和衰老。具体来说，翻译机械的核心部件以及P型车身相互作用组件加速了老化过程。翻译启动的扰动会加速老化。因此，通过 FRAP 测量全球蛋白质合成速率是衰老过程的直接读出。

Research

JoVE Journal

Biology

Assessment of de novo Protein Synthesis Rates in Caenorhabditis elegans

Osmotic Avoidance in Caenorhabditis elegans: Synaptic Function of Two Genes, Orthologues of Human NRXN1 and NLGN1, as Candidates for Autism

Osmotic Avoidance in Caenorhabditis elegans: Synaptic Function of Two Genes, Orthologues of Human NRXN1 and NLGN1, as Candidates for Autism

渗透避免IN线虫：突触功能的两个基因，人类同源 NRXN1 NLGN1，作为候选人自闭症

Neurexins and neuroligins are cell adhesion molecules present in excitatory and inhibitory synapses, and they are required for correct neuron network ...

科研

生物学

Time-lapse Microscopy of Early Embryogenesis in Caenorhabditis elegans

Time-lapse Microscopy of Early Embryogenesis in Caenorhabditis elegans

时间推移显微镜早期胚胎发育线虫

Caenorhabditis elegans has often been used as a model system in studies of early developmental processes.  The transparency of the 
embryos, the genetic ...

Basic Caenorhabditis elegans Methods: Synchronization and Observation

Basic Caenorhabditis elegans Methods: Synchronization and Observation

基本的<em&gt;线虫</em&gt;方法：同步和观察

Research into the molecular and developmental biology of the nematode Caenorhabditis elegans was begun in the early seventies by Sydney Brenner and it has ...

Biochemical and High Throughput Microscopic Assessment of Fat Mass in Caenorhabditis Elegans

Biochemical and High Throughput Microscopic Assessment of Fat Mass in Caenorhabditis Elegans

生化和高通量的微观评估的脂肪含量<em&gt;秀丽隐杆线虫</em

The nematode C. elegans has emerged as an important  model for the study of conserved genetic pathways regulating fat metabolism as  it relates to human ...

Prostaglandin Extraction and Analysis in Caenorhabditis elegans

Prostaglandin Extraction and Analysis in Caenorhabditis elegans

前列腺素的提取与分析<em&gt;线虫</em

Caenorhabditis elegans is emerging as a powerful animal model to study the biology of lipids1-9. Prostaglandins are an important class of eicosanoids, ...

Protein WISDOM: A Workbench for In silico De novo Design of BioMolecules

Protein WISDOM: A Workbench for In silico De novo Design of BioMolecules

蛋白质智慧：一个工作台<em&gt;硅片</em&gt;<em&gt;从头</em&gt;生物分子设计

The aim of de novo protein design is to find the amino acid sequences that will fold into a desired 3-dimensional structure with improvements in specific ...

Using Caenorhabditis elegans as a Model System to Study Protein Homeostasis in a Multicellular Organism

Using Caenorhabditis elegans as a Model System to Study Protein Homeostasis in a Multicellular Organism

The folding and assembly of proteins is essential for protein function, the long-term health of the cell, and longevity of the organism. Historically, the ...

Methods to Study Changes in Inherent Protein Aggregation with Age in Caenorhabditis elegans

Methods to Study Changes in Inherent Protein Aggregation with Age in Caenorhabditis elegans

方法研究线虫线虫中固有的蛋白质聚集与年龄的变化

In the last decades, the prevalence of neurodegenerative disorders, such as Alzheimer's disease (AD) and Parkinson's disease (PD), has grown. These ...

De Novo Generation of Somatic Stem Cells by YAP/TAZ

De Novo Generation of Somatic Stem Cells by YAP/TAZ

从头用狂吠/狂暴法生成体细胞干细胞

Here we present protocols to isolate primary differentiated cells and turn them into stem/progenitor cells (SCs) of the same lineage by transient ...

Detecting Protein Subcellular Localization by Green Fluorescence Protein Tagging and 4',6-Diamidino-2-phenylindole Staining in Caenorhabditis elegans

Detecting Protein Subcellular Localization by Green Fluorescence Protein Tagging and 4',6-Diamidino-2-phenylindole Staining in Caenorhabditis elegans

秀丽线虫绿色荧光蛋白标记和 4 '、6-Diamidino-2-苯基吲哚染色检测蛋白质亚细胞定位

In this protocol, a green fluorescence protein (GFP) fusion protein and 4',6-diamidino-2-phenylindole (DAPI) staining are used to track protein ...

卡诺哈布迪蒂斯 伊利根斯新蛋白合成率评估

卡诺哈布迪蒂斯伊利根斯新蛋白合成率评估