作者: Landlocked 時(shí)間: 2025-3-21 23:47 作者: Pigeon 時(shí)間: 2025-3-22 01:33
Purification of Bacterial RNA Polymerase: Tools and Protocols, the use of multicistronic vectors for expression of the . enzyme in large quantities and in a highly active form. Here we describe the commonly used vectors and procedures for purification of the . RNA polymerase.作者: flaggy 時(shí)間: 2025-3-22 05:02
Differential- und Integralrechnung the use of multicistronic vectors for expression of the . enzyme in large quantities and in a highly active form. Here we describe the commonly used vectors and procedures for purification of the . RNA polymerase.作者: neolith 時(shí)間: 2025-3-22 11:43 作者: EXALT 時(shí)間: 2025-3-22 16:55 作者: 痛苦一下 時(shí)間: 2025-3-22 18:25
In Situ Footprinting of , Transcription Elongation Complex with Chloroacetaldehyde,cetaldehyde for in situ footprinting of . elongation complex temporarily halted by a protein roadblock. The method provides valuable information on the dynamic features of transcriptionally engaged RNA polymerase within the cellular environment.作者: SMART 時(shí)間: 2025-3-23 00:50
Using Solutes and Kinetics to Probe Large Conformational Changes in the Steps of Transcription Initnding assays from series used to determine the urea dependence of open complex formation and dissociation with . RNA polymerase and phage λP. promoter DNA. Then, we describe the subsequent data analysis and interpretation of these solute effects.作者: Merited 時(shí)間: 2025-3-23 05:09 作者: Herbivorous 時(shí)間: 2025-3-23 08:37 作者: 脆弱帶來(lái) 時(shí)間: 2025-3-23 12:02
https://doi.org/10.1007/978-3-8348-9245-4e sequence and phosphorylation status of the 5′ ends of RNAs. 5′ specific RNA-seq can be used to analyze transcription initiation and posttranscriptional processing of RNAs with single base pair resolution on a genome-wide level.作者: 侵害 時(shí)間: 2025-3-23 16:42
Wilfried Pla?mann,Detlef Schulzcetaldehyde for in situ footprinting of . elongation complex temporarily halted by a protein roadblock. The method provides valuable information on the dynamic features of transcriptionally engaged RNA polymerase within the cellular environment.作者: Custodian 時(shí)間: 2025-3-23 18:21
https://doi.org/10.1007/978-3-8348-9245-4nding assays from series used to determine the urea dependence of open complex formation and dissociation with . RNA polymerase and phage λP. promoter DNA. Then, we describe the subsequent data analysis and interpretation of these solute effects.作者: 冷峻 時(shí)間: 2025-3-23 22:18
https://doi.org/10.1007/978-3-8348-9245-4(1) single-round transcription, (2) walking of RNAP to any defined template position, and (3) discrimination of transcripts that are associated with RNAP from those that are released to solution. This methodology is based on untagged proteins transcribing biotin- and digoxigenin-labeled DNA templates in association with paramagnetic particles.作者: 廢墟 時(shí)間: 2025-3-24 03:15 作者: Invigorate 時(shí)間: 2025-3-24 07:13
Differential- und Integralrechnungstrand designs where translocation of RNA polymerase from a pre-translocation to a post-translocation state results in disruption of stacking interactions of fluorophore with neighboring bases, with a concomitant large increase in fluorescence intensity.作者: 使絕緣 時(shí)間: 2025-3-24 13:49
https://doi.org/10.1007/978-3-8348-9245-4through its stepwise translocation towards RNA polymerase. This system can be used to study the effects of concurrent translation on RNA chain elongation and to elucidate the interface between the two macromolecular complexes.作者: 水汽 時(shí)間: 2025-3-24 16:39 作者: 多山 時(shí)間: 2025-3-24 19:34
https://doi.org/10.1007/978-3-8348-9245-4 easily expressed and purified from heterologous expression systems. Forming functional RNA polymerase involves simply combining the different subunits under denaturing conditions and slowly removing the denaturant.作者: 我沒(méi)有命令 時(shí)間: 2025-3-25 01:27 作者: 合同 時(shí)間: 2025-3-25 06:31 作者: Nmda-Receptor 時(shí)間: 2025-3-25 07:48
Monitoring Translocation of Multisubunit RNA Polymerase Along the DNA with Fluorescent Base Analogustrand designs where translocation of RNA polymerase from a pre-translocation to a post-translocation state results in disruption of stacking interactions of fluorophore with neighboring bases, with a concomitant large increase in fluorescence intensity.作者: 打火石 時(shí)間: 2025-3-25 12:02
Methods for the Assembly and Analysis of In Vitro Transcription-Coupled-to-Translation Systems,through its stepwise translocation towards RNA polymerase. This system can be used to study the effects of concurrent translation on RNA chain elongation and to elucidate the interface between the two macromolecular complexes.作者: Constrain 時(shí)間: 2025-3-25 16:26 作者: 古老 時(shí)間: 2025-3-25 20:20
Transcription in Archaea: Preparation of , Transcription Machinery, easily expressed and purified from heterologous expression systems. Forming functional RNA polymerase involves simply combining the different subunits under denaturing conditions and slowly removing the denaturant.作者: 不遵守 時(shí)間: 2025-3-26 03:06 作者: Explicate 時(shí)間: 2025-3-26 05:08
1064-3745 ation advice from the experts.This volume. .is designed to be a resource of proven techniques and approaches for probing the activities of bacterial, eukaryotic, and archaeal RNA polymerases. This book features a collection of .in vitro .and .in vivo. technologies that will permit researchers to pur作者: idiopathic 時(shí)間: 2025-3-26 11:27 作者: 狗窩 時(shí)間: 2025-3-26 13:35 作者: 盡管 時(shí)間: 2025-3-26 19:14
Wilfried Pla?mann,Detlef Schulzassemble a functional . enzyme. Standard in vitro transcription assays can be used to examine the different stages of transcription. In this chapter, we describe how some of these assays have been optimized for . RNA polymerase, which transcribes at much higher temperatures than many other transcription complexes.作者: myriad 時(shí)間: 2025-3-26 23:54 作者: 話 時(shí)間: 2025-3-27 02:55
Direct Competition Assay for Transcription Fidelity,ve analyses of transcription fidelity, applicable to analyses of RNA polymerase selectivity against misincorporation, incorporation of dNMPs, and chemically modified rNMP analogues. The method is based on different electrophoretic mobility of RNA oligomers of the same length but differing in sequence.作者: epinephrine 時(shí)間: 2025-3-27 05:29
Transcription in Archaea: In Vitro Transcription Assays for mjRNAP,assemble a functional . enzyme. Standard in vitro transcription assays can be used to examine the different stages of transcription. In this chapter, we describe how some of these assays have been optimized for . RNA polymerase, which transcribes at much higher temperatures than many other transcription complexes.作者: 座右銘 時(shí)間: 2025-3-27 11:02 作者: 地名詞典 時(shí)間: 2025-3-27 14:26
Wilfried Pla?mann,Detlef Schulztive RNA bands detected on gels reflects the molar ratios and quantities of each RNA product, regardless of length. The radiolabeled RNAs are isolated by hybridization to a biotinylated oligonucleotide and captured on magnetic beads. We also describe the use of antibodies to investigate mechanistic aspects of promoter proximal pausing.作者: Root494 時(shí)間: 2025-3-27 20:01
https://doi.org/10.1007/978-3-8348-9245-4 integral subunits (Arimbasseri et al. Transcription 4(6), 2013). This makes pol III a valuable model for dissecting intrinsic molecular mechanisms of eukaryotic transcription termination. In this chapter, we provide protocols we adapted to study the biochemistry of transcription termination by . pol III.作者: 舊式步槍 時(shí)間: 2025-3-27 22:00 作者: 良心 時(shí)間: 2025-3-28 05:03 作者: Left-Atrium 時(shí)間: 2025-3-28 09:21 作者: 滑動(dòng) 時(shí)間: 2025-3-28 14:12
Biochemical Analysis of Transcription Termination by RNA Polymerase III from Yeast ,, integral subunits (Arimbasseri et al. Transcription 4(6), 2013). This makes pol III a valuable model for dissecting intrinsic molecular mechanisms of eukaryotic transcription termination. In this chapter, we provide protocols we adapted to study the biochemistry of transcription termination by . pol III.作者: 珠寶 時(shí)間: 2025-3-28 18:30
Purification of Active RNA Polymerase I from Yeast, features of an enzyme, in vitro assays are critical. Here we describe a method for purifying RNA polymerase I. This approach yields enzyme of sufficiently high quantity and activity for an array of experiments directed at describing the enzymatic properties of RNA polymerase I in detail.作者: agenda 時(shí)間: 2025-3-28 19:44
Book 2015polymerases. This book features a collection of .in vitro .and .in vivo. technologies that will permit researchers to purify and probe the position and stability of RNA polymerase complexes at different points of the transcription cycle, analyze the various translocations and intermolecular movement作者: 幻想 時(shí)間: 2025-3-28 23:56
Irina Artsimovitch,Thomas J. SantangeloIncludes cutting-edge methods and protocols.Provides step-by-step detail essential for reproducible results.Contains key notes and implementation advice from the experts作者: decode 時(shí)間: 2025-3-29 05:10 作者: doxazosin 時(shí)間: 2025-3-29 08:50
Bacterial Transcriptional Control978-1-4939-2392-2Series ISSN 1064-3745 Series E-ISSN 1940-6029 作者: Fantasy 時(shí)間: 2025-3-29 12:55
Wilfried Pla?mann,Detlef Schulz promoters, induce pausing during RNA chain synthesis, and control transcription termination. These interactions are dissected using footprinting assays, in which a bound protein protects nucleic acids from the digestion by nucleases or modification by chemical probes. Exonuclease III is frequently 作者: forbid 時(shí)間: 2025-3-29 18:20
Differential- und Integralrechnungnal apparatus is by far the best characterized, with numerous RNA polymerase mutants and auxiliary factors isolated and analyzed in great detail. Since the . enzyme was refractory to crystallization, structural studies have been focused on . RNA polymerases, revealing atomic details of the catalytic作者: 大方一點(diǎn) 時(shí)間: 2025-3-29 21:15
Differential- und Integralrechnunge. In a stopped-flow setup, this method is capable of resolving a single base-pair translocation motion of RNA polymerase in real time. In a conventional spectrofluorometer, this method can be employed for studies of the time-averaged distribution of RNA polymerase on the DNA template. This method u作者: Nefarious 時(shí)間: 2025-3-30 00:57 作者: 敵意 時(shí)間: 2025-3-30 07:01 作者: jocular 時(shí)間: 2025-3-30 10:18 作者: Affiliation 時(shí)間: 2025-3-30 12:42 作者: separate 時(shí)間: 2025-3-30 17:36
https://doi.org/10.1007/978-3-8348-9245-4 polymerases are currently elucidated by structural, genetic, and biochemical approaches. Here, we describe a fast and reliable approach to quantitative analyses of transcription fidelity, applicable to analyses of RNA polymerase selectivity against misincorporation, incorporation of dNMPs, and chem作者: 輕快帶來(lái)危險(xiǎn) 時(shí)間: 2025-3-30 22:59
https://doi.org/10.1007/978-3-8348-9245-4uct, nucleotide substrates, metal cofactors, and regulatory molecules that bind to distinct RNAP sites to modulate its activity. RNAP is also inhibited by several known antibiotics and is a promising target for development of novel antibacterial compounds. Despite great progress in structural analys作者: Adulate 時(shí)間: 2025-3-31 04:23
https://doi.org/10.1007/978-3-8348-9245-4mpared to its pol II counterpart, Pol III has several advantages, including the relative simplicity, stability, and more direct connectivity of its transcription machinery. Only two transcription factor complexes, TFIIIB and TFIIIC, are required to faithfully initiate and direct multiple rounds of t作者: 休閑 時(shí)間: 2025-3-31 05:04
Wilfried Pla?mann,Detlef Schulzlymerase–promoter complex proceeds through multiple intermediates. Short promoter fragments can be used as a tool to dissect RNA polymerase–promoter interactions and to pinpoint elements responsible for specific properties of the entire promoter complex. A recently developed fluorometric molecular b作者: 過(guò)于平凡 時(shí)間: 2025-3-31 13:01
https://doi.org/10.1007/978-3-8348-9245-4RNA (5′ specific RNA-seq). The protocol describes how cDNA libraries for 5′ specific RNA-seq can be tailored to analyze specific classes of RNAs based upon the phosphorylation status of the 5′ end. Thus, the analysis of cDNA libraries generated by these methods provides information regarding both th作者: 籠子 時(shí)間: 2025-3-31 14:37 作者: 隨意 時(shí)間: 2025-3-31 21:04 作者: legitimate 時(shí)間: 2025-3-31 23:35
https://doi.org/10.1007/978-3-8348-9245-4n include pausing, backtracking, arrest, and transcription termination, and it is critical to determine whether the absence of continued synthesis is transient or permanent. Here we describe mechanisms to generate large quantities of stable archaeal elongation complexes on a solid support to permit 作者: facetious 時(shí)間: 2025-4-1 03:37 作者: 沉積物 時(shí)間: 2025-4-1 09:54 作者: amyloid 時(shí)間: 2025-4-1 10:51 作者: 溫和女孩 時(shí)間: 2025-4-1 16:05
https://doi.org/10.1007/978-3-322-80856-1g transcription. Here, we describe a highly purified experimental system that recapitulates many important properties of transcribed chromatin and the key aspects of hFACT action during this process in vitro. We present the protocols describing how to prepare different forms of nucleosomes, includin
