Abstract:
:Dicer is a multidomain ribonuclease that processes double-stranded RNAs (dsRNAs) to 21-nt small interfering RNAs (siRNAs) during RNA interference and excises microRNAs (miRNAs) from precursor hairpins. PAZ and PIWI domain (PPD) proteins, also involved in RNAi and miRNA function, are the best-characterized proteins known to interact with Dicer. PPD proteins are the core constituents of effector complexes, RISCs and miRNPs, mediating siRNA and miRNA function. In this chapter we describe overexpression and purification of recombinant human Dicer, its biochemical properties, and mapping of domains responsible for Dicer-PPD protein interactions.
journal_name
Methods Enzymoljournal_title
Methods in enzymologyauthors
Kolb FA,Zhang H,Jaronczyk K,Tahbaz N,Hobman TC,Filipowicz Wdoi
10.1016/S0076-6879(04)92019-8subject
Has Abstractpub_date
2005-01-01 00:00:00pages
316-36eissn
0076-6879issn
1557-7988pii
S0076687904920198journal_volume
392pub_type
杂志文章abstract::The combined use of rotating-frame relaxation methods, temperature-dependent measurements of line shapes and magnetization transfer experiments allows in favorable cases the examination in some detail of exchange processes that occur on the millisecond time scale. It is possible to determine not only the rate constant...
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doi:10.1016/S0076-6879(07)38024-5
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abstract::Inducible expression systems represent the founding technology for the emergence of synthetic biology in mammalian cells. The core molecules in these systems are bacterial regulator proteins that bind to or dissociate from a cognate DNA operator sequence in response to an exogenous stimulus like a small-molecule induc...
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doi:10.1016/B978-0-12-385075-1.00011-1
更新日期:2011-01-01 00:00:00
abstract::Staphylococcus aureus ribonuclease III (Sa-RNase III) belongs to the enzyme family known to process double-stranded RNAs consisting of two turns of the RNA helix. Although the enzyme is thought to play a role in ribosomal RNA processing and gene regulation, the deletion of the rnc gene in S. aureus does not affect cel...
journal_title:Methods in enzymology
pub_type: 杂志文章
doi:10.1016/S0076-6879(08)02216-7
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pub_type: 杂志文章,评审
doi:10.1016/bs.mie.2019.06.016
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journal_title:Methods in enzymology
pub_type: 杂志文章
doi:10.1016/s0076-6879(02)45012-4
更新日期:2002-01-01 00:00:00
abstract::Scanning transmission electron microscopy (STEM) of unstained, freeze-dried biological macromolecules in the dark-field mode provides an image based on the number of electrons elastically scattered by the constituent atoms of the macromolecule. The image of each isolated particle provides information about the project...
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doi:10.1016/S0076-6879(08)36027-3
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abstract::We present an in vitro method to measure how Rab4 and other regulatory proteins affect microtubule-based organelle motility. The protocols utilize small-volume, disposable "microchambers" designed for epifluorescence, confocal, or other microscope platforms and into which microtubules, organelles, and primary and fluo...
journal_title:Methods in enzymology
pub_type: 杂志文章
doi:10.1016/S0076-6879(05)03009-0
更新日期:2005-01-01 00:00:00
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doi:10.1016/S0076-6879(10)75010-2
更新日期:2010-01-01 00:00:00
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doi:10.1016/bs.mie.2016.09.037
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journal_title:Methods in enzymology
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doi:10.1016/B978-0-12-381270-4.00001-9
更新日期:2011-01-01 00:00:00
abstract::Epitope tagging has provided a useful experimental strategy with widespread applicability. The ample variety of epitope tag systems that have been put to use to date provide a collection of attributes relevant to virtually any experimental system. As a consequence, epitope tagging will continue to be a valuable tool f...
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abstract::Sulfatases hydrolyze sulfate esters on a variety of molecules including glycosaminoglycans, sulfoglycolipids, and cytosolic steroids. These enzymes are found in a wide range of organisms with their basic enzymatic mechanisms broadly conserved. In mammals, many of the sulfatases localize in the lysosome and exhibit enz...
journal_title:Methods in enzymology
pub_type: 杂志文章
doi:10.1016/S0076-6879(06)16015-2
更新日期:2006-01-01 00:00:00
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journal_title:Methods in enzymology
pub_type: 杂志文章
doi:10.1016/S0076-6879(07)00429-6
更新日期:2008-01-01 00:00:00
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pub_type: 杂志文章
doi:10.1016/bs.mie.2019.05.058
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abstract::Hsp70 chaperones and their obligatory J-protein cochaperones function together in many cellular processes. Via cycles of binding to short stretches of exposed amino acids on substrate proteins, Hsp70/J-protein chaperones not only facilitate protein folding but also drive intracellular protein transport, biogenesis of ...
journal_title:Methods in enzymology
pub_type: 杂志文章,评审
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pub_type: 杂志文章,评审
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更新日期:2008-01-01 00:00:00
abstract::Ubiquitin-like protein (Ubl) ligation is common to diverse archaea and targets many cellular pathways, including those associated with sulfur mobilization, and also tags proteins as substrates for degradation by the proteasome. Here we highlight protocols to assay proteasome function and Ubl ligation in archaea. A cha...
journal_title:Methods in enzymology
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journal_title:Methods in enzymology
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