Abstract:
:The kinetics of PaeR7 endonuclease-catalysed cleavage reactions of fluorophor-labeled oligonucleotide substrates have been examined using fluorescence resonance energy transfer (FRET). A series of duplex substrates were synthesized with an internal CTCGAG PaeR7 recognition site and donor (fluorescein) and acceptor (rhodamine) dyes conjugated to the opposing 5' termini. The time-dependent increase in donor fluorescence resulting from restriction cleavage of these substrates was continuously monitored and the initial rate data was fitted to the Michaelis-Menten equation. The steady state kinetic parameters for these substrates were in agreement with the rate constants obtained from a gel electrophoresis-based fixed time point assay using radiolabeled substrates. The FRET method provides a rapid continuous assay as well as high sensitivity and reproducibility. These features should make the technique useful for the study of DNA-cleaving enzymes.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Ghosh SS,Eis PS,Blumeyer K,Fearon K,Millar DPdoi
10.1093/nar/22.15.3155subject
Has Abstractpub_date
1994-08-11 00:00:00pages
3155-9issue
15eissn
0305-1048issn
1362-4962journal_volume
22pub_type
杂志文章abstract::A simple, two-step method is described for the synthesis of the 5'-pyro- and triphosphate derivatives of 3'-5' ApA, ApG, GpA and GpG. The readily accessible 2'(3')-5' ApA, ApG, GpA and GpG were converted in one step to the corresponding 5'-phosphoramidate derivatives which were then transformed to the 5'-pyro- and tri...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/5.8.2945
更新日期:1978-08-01 00:00:00
abstract::Multivalent cations are known to condense DNA into higher ordered structures, including toroids and rods. Here we report that solid supports treated with monovalent or multivalent cationic silanes, followed by removal of soluble molecules, can condense DNA. The mechanism of this surface-directed condensation depends o...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.2.588
更新日期:1998-01-15 00:00:00
abstract::Selected groups of isolated 14C-labelled proteins from E. coli 30S ribosomal subunits were reconstituted with 32P-labelled 16S RNA, and the reconstituted complexes were partially digested with ribonuclease A. RNA fragments protected by the proteins were separated by gel electrophoresis and subjected to sequence analys...
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pub_type: 杂志文章
doi:10.1093/nar/16.4.1233
更新日期:1988-02-25 00:00:00
abstract::Robust methods to detect DNA-binding proteins from structures of unknown function are important for structural biology. This paper describes a method for identifying such proteins that (i) have a solvent accessible structural motif necessary for DNA-binding and (ii) a positive electrostatic potential in the region of ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh803
更新日期:2004-09-08 00:00:00
abstract::Recent studies have shown that homologous recombination (HR) requires chromatin repression as well as relaxation at DNA double strand breaks (DSBs). HP1 and SUV39H1/2 are repressive factors essential for HR. Here, we identify SETDB1 as an additional compacting factor promoting HR. Depletion of HP1, SUV39, SETDB1 or BR...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv722
更新日期:2015-09-18 00:00:00
abstract::DNA strand exchange, the central step of homologous recombination, is considered to occur approximately independently of DNA sequence content. However, certain prokaryotic and eukaryotic genomic loci display either an enhanced or reduced frequency of genetic exchange. Here we show that the Homo sapiens DNA strand exch...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkl355
更新日期:2006-05-24 00:00:00
abstract::Calf lens messenger fractions have been purified on poly(dT)-cellulose. The purified RNA's have been analyzed as nucleosides by a method which can be used on a micro-scale with highly reproduceble results. Both the 10S and 14S lens mRNA contain poly A sequences. The poly A content found in 14S mRNA cannot account for ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/1.2.245
更新日期:1974-02-01 00:00:00
abstract::OGG1 initiated base excision repair (BER) is the major pathway for repair of oxidative DNA base damage 8-oxoguanine (8-oxoG). Here, we report that RECQL4 DNA helicase, deficient in the cancer-prone and premature aging Rothmund-Thomson syndrome, physically and functionally interacts with OGG1. RECQL4 promotes catalytic...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa392
更新日期:2020-07-09 00:00:00
abstract::Modified bases, such as O6-methylguanines, are produced in cells exposed to alkylating agents and cause apoptosis. In human cells treated with N-methyl-N-nitrosourea, we detected a protein complex composed of MutSalpha, MutLalpha and PCNA on damaged DNA by immunoprecipitation method using chromatin extracts, in which ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gki878
更新日期:2005-10-04 00:00:00
abstract::To examine the influence of thyrotropin (TSH) on the thyroglobulin (Tgb) mRNA content, the latter was evaluated in the cytoplasm of hog thyroid cells cultured in the absence (control cells) or presence of TSH. The Tgb mRNA levels were determined by, (i) kinetics of hybridization to sheep Tgb cDNA, (ii) capacity of cod...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/6.10.3353
更新日期:1979-07-25 00:00:00
abstract::The beta protein encoded by the Streptococcus pyogenes plasmid pSM19035 is a site-specific recombinase involved in both resolution of plasmid multimers into monomers and DNA inversion. It has been proposed that the DNA region to which the beta recombinase binds to mediate recombination includes a promoter from which o...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/22.10.1855
更新日期:1994-05-25 00:00:00
abstract::Light is a major external factor in regulating seed germination. Photoreceptor phytochrome B (PHYB) plays a predominant role in promoting seed germination in the initial phase after imbibition, partially by repressing phytochrome-interacting factor1 (PIF1). However, the mechanism underlying the PHYB-PIF1-mediated tran...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx283
更新日期:2017-07-07 00:00:00
abstract::A novel method for sequence specific double strand DNA cleavage using PNA (peptide nucleic acid) targeting is described. Nuclease S1 digestion of double stranded DNA gives rise to double strand cleavage at an occupied PNA strand displacement binding site, and under optimized conditions complete cleavage can be obtaine...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/21.9.2103
更新日期:1993-05-11 00:00:00
abstract::In a gene targeting experiment, the generation of a targeting construct often requires complex DNA manipulations. We developed a set of cassettes and plasmids useful for creating targeting vectors to modify the mammalian genome. A positive selection marker cassette (PGK/EM7p-npt), which included dual prokaryotic and e...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gni055
更新日期:2005-03-22 00:00:00
abstract::The binding of nine aminoalkyl adenylates to isoleucyl-tRNA synthetase from Escherichia coli MRE 600 was measured and compared with the binding of the cognate amino acids. It was found that they bind rather tightly to the enzyme, the Kd's ranging from 3.1.10(-4) M with glycinol-AMP ester to 3.7.10(-9) M with L-isoleuc...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/4.3.673
更新日期:1977-03-01 00:00:00
abstract::For over 10 years, Binding MOAD (Mother of All Databases; http://www.BindingMOAD.org) has been one of the largest resources for high-quality protein-ligand complexes and associated binding affinity data. Binding MOAD has grown at the rate of 1994 complexes per year, on average. Currently, it contains 23,269 complexes ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gku1088
更新日期:2015-01-01 00:00:00
abstract::The reactivities of the two cysteine thiol groups of calf thymus F3 histone were investigated using 5,5'-dithiobis-[2- nitrobenzoic acid], (DTNB). In isolated histone, both thiol groups were available for reaction. However, analysis of reaction profiles of native deoxyribonucleohistone, (DNH), in various solvent condi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/1.2.203
更新日期:1974-02-01 00:00:00
abstract::Bacterial small regulatory RNAs (sRNAs) play a major role in the regulation of various cellular functions. Most sRNAs interact with mRNA targets via an antisense mechanism, modifying their translation and/or degradation. Despite considerable progresses in discovering sRNAs in Gram-positive bacteria, their functions, f...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx190
更新日期:2017-05-05 00:00:00
abstract::Synthetic DNA fragments were constructed to determine the effect of G tracts, in conjunction with periodically spaced A tracts, on DNA bends. Relative length measurements showed that the G tracts spaced at the half helical turn enhanced the DNA bend. When the G tract was interrupted with a thymine or shortened to one ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.4.817
更新日期:1990-02-25 00:00:00
abstract::TAP/hNXF1 is a key factor that mediates general cellular mRNA export from the nucleus, and its orthologs are structurally and functionally conserved from yeast to humans. Metazoans encode additional proteins that share homology and domain organization with TAP/hNXF1, suggesting their participation in mRNA metabolism; ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gki706
更新日期:2005-07-13 00:00:00
abstract::The mouse is widely used as system to study human genetic mechanisms. However, extensive rewiring of transcriptional regulatory networks often confounds translation of findings between human and mouse. Site-specific gain and loss of individual transcription factor binding sites (TFBS) has caused functional divergence ...
journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:2018-02-28 00:00:00
abstract::The Protein Circular Dichroism Data Bank (PCDDB) has been in operation for more than 5 years as a public repository for archiving circular dichroism spectroscopic data and associated bioinformatics and experimental metadata. Since its inception, many improvements and new developments have been made in data display, se...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw796
更新日期:2017-01-04 00:00:00
abstract::The active 3D conformation of the spliceosome's catalytic U2/U6 RNA core is stabilised by a network of secondary and tertiary RNA interactions, but also depends on spliceosomal proteins for its formation. To determine the contribution towards splicing of specific RNA secondary and tertiary interactions in the U2/U6 RN...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky966
更新日期:2018-12-14 00:00:00
abstract::Two different genomic genes for the COUP-transcription factor, COUP-TF I and COUP-TF II, have been isolated from a human cosmid genomic library using a [32P]-labeled cDNA probe. Data obtained from Southern blot analysis of these cosmid clones indicated that two closely related genes exist in the human genome and have ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.23.6857
更新日期:1990-12-11 00:00:00
abstract::Computational analysis of biosynthetic gene clusters (BGCs) has revolutionized natural product discovery by enabling the rapid investigation of secondary metabolic potential within microbial genome sequences. Grouping homologous BGCs into Gene Cluster Families (GCFs) facilitates mapping their architectural and taxonom...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa812
更新日期:2021-01-08 00:00:00
abstract::PAHdb (http://www.mcgill.ca/pahdb ) is a curated relational database (Fig. 1) of nucleotide variation in the human PAH cDNA (GenBank U49897). Among 328 different mutations by state (Fig. 2) the majority are rare mutations causing hyperphenylalaninemia (HPA) (OMIM 261600), the remainder are polymorphic variants without...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.1.220
更新日期:1998-01-01 00:00:00
abstract::Polynucleotides containing adenosine and 8-azidoadenosine or inosine and 8-azidoinosine residues have been prepared from mixtures of nucleoside diphosphates using polynucleotide phosphorylase from Escherichia coli. These copolymers can form complexes with polyuridylic or polycytidylic acids respectively. Single strand...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/8.7.1675
更新日期:1980-04-11 00:00:00
abstract::Bacterial small RNAs (sRNAs) are a heterogeneous group of post-transcriptional regulators that often act at the heart of large networks. Hundreds of sRNAs have been discovered by genome-wide screens and most of these sRNAs exert their functions by base-pairing with target mRNAs. However, studies addressing the molecul...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky765
更新日期:2018-11-16 00:00:00
abstract::Solution structures of DNA duplexes containing oxanine (Oxa, O) opposite a cytosine (O:C duplex) and opposite a thymine (O:T duplex) have been solved by the combined use of (1)H NMR and restrained molecular dynamics calculation. One mismatch pair was introduced into the center of the 11-mer duplex of [d(GTGACO(6)CACTG...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr872
更新日期:2012-02-01 00:00:00
abstract::Transgenic mice suitable for the in vivo assay of suspected mutagens at the chromosome level have been constructed by stable integration of a lambda phage shuttle vector. The shuttle vector, which contains a beta-galactosidase (beta-gal) target gene, can be rescued from genomic DNA with in vitro packaging extracts. Mu...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.10.3007
更新日期:1990-05-25 00:00:00