Abstract:
:Labeling of a covalently closed circular double-stranded DNA was achieved using a so-called 'padlock oligonucleotide'. The oligonucleotide was targeted to a sequence which is present in the replication origin of phage f1 and thus in numerous commonly used plasmids. After winding around the double-stranded target DNA sequence by ligand-induced triple helix formation, a biotinylated oligonucleotide was circularized using T4 DNA ligase and in this way became catenated to the plasmid. A gel shift assay was developed to measure the extent of plasmid modification by the padlock oligonucleotide. A similar assay showed that a modified supercoiled plasmid was capable of binding one streptavidin molecule thanks to the biotinylated oligonucleotide and that this binding was quantitative. The catenated complex was visualized by electron and atomic force microscopies using streptavidin conjugates or single strand-binding proteins as protein tags for the padlock oligonucleotide. This method provides a versatile tool for plasmid functionalization which offers new perspectives in the physical study of supercoiled DNA and in the development of improved vectors for gene therapy.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Roulon T,Coulaud D,Delain E,Le Cam E,Hélène C,Escudé Cdoi
10.1093/nar/30.3.e12keywords:
subject
Has Abstractpub_date
2002-02-01 00:00:00pages
E12issue
3eissn
0305-1048issn
1362-4962journal_volume
30pub_type
杂志文章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::A sensitive gel retention assay has been utilized to detect proteins from uninfected Hela nuclei which interact with the adenovirus type 2 enhancer. This assay has been employed to monitor fractionation of nuclear extracts. Three enhancer binding factors were resolved by chromatography on DEAE-Sepharose and one of the...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/15.6.2719
更新日期:1987-03-25 00:00:00
abstract::The MyHits web site (http://myhits.isb-sib.ch) is an integrated service dedicated to the analysis of protein sequences. Since its first description in 2004, both the user interface and the back end of the server were improved. A number of tools (e.g. MAFFT, Jacop, Dotlet, Jalview, ESTScan) were added or updated to imp...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkm352
更新日期:2007-07-01 00:00:00
abstract::The wobble uridine of certain bacterial and mitochondrial tRNAs is modified, at position 5, through an unknown reaction pathway that utilizes the evolutionarily conserved MnmE and GidA proteins. The resulting modification (a methyluridine derivative) plays a critical role in decoding NNG/A codons and reading frame mai...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkp762
更新日期:2009-11-01 00:00:00
abstract::DNA labeled for 15 minutes during UV induced repair synthesis is two-fold more sensitive to micrococcal nuclease than the bulk nuclear DNA. As the length of the labeling period increases from 15 minutes to 4 hours the nuclease sensitivity of repair labeled DNA approaches that of bulk chromatin. Pulse-chase experiments...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/9.1.203
更新日期:1981-01-10 00:00:00
abstract::Alternative splicing is widespread in mammalian gene expression, and variant splice patterns are often specific to different stages of development, particular tissues or a disease state. There is a need to systematically collect data on alternatively spliced exons, introns and splice isoforms, and to annotate this dat...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh030
更新日期:2004-01-01 00:00:00
abstract::The PathoSystems Resource Integration Center (PATRIC) is the bacterial Bioinformatics Resource Center funded by the National Institute of Allergy and Infectious Diseases (https://www.patricbrc.org). PATRIC supports bioinformatic analyses of all bacteria with a special emphasis on pathogens, offering a rich comparative...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkz943
更新日期:2020-01-08 00:00:00
abstract::MicroRNAs (miRNAs) are approximately 22 nucleotide (nt) long and play important roles in post-transcriptional regulation in both plants and animals. In animals, precursor (pre-) miRNAs are ∼70 nt hairpins produced by Drosha cleavage of long primary (pri-) miRNAs in the nucleus. Exportin-5 (XPO5) transports pre-miRNAs ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky306
更新日期:2018-06-20 00:00:00
abstract::Bacterial tyrosine-kinases share no resemblance with their eukaryotic counterparts and they have been unified in a new protein family named BY-kinases. These enzymes have been shown to control several biological functions in the bacterial cells. In recent years biochemical studies, sequence analyses and structure reso...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr915
更新日期:2012-01-01 00:00:00
abstract::The solution structures of two 27 nt RNA hairpins and their complexes with cobalt(III)-hexammine [Co(NH(3))(6)(3+)] were determined by NMR spectroscopy. The RNA hairpins are variants of the P4 region from Escherichia coli RNase P RNA: a U-to-A mutant changing the identity of the bulged nucleotide, and a U-to-C, C-to-U...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh961
更新日期:2004-12-02 00:00:00
abstract::Eukaryotic cells possess several distinct mismatch repair pathways. A mismatch can be introduced in retroviral double-stranded DNA by a pre-existing mutation within the primer binding site (PBS) of the viral RNA genome. In order to evaluate mismatch repair of retroviral double-stranded DNA, Moloney leukemia virus (MLV...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/28.12.2302
更新日期:2000-06-15 00:00:00
abstract::Tight cell-cycle regulation of the histone H4-K20 methyltransferase PR-Set7 is essential for the maintenance of genome integrity. In mammals, this mainly involves the interaction of PR-Set7 with the replication factor PCNA, which triggers the degradation of the enzyme by the CRL4CDT2 E3 ubiquitin ligase. PR-Set7 is al...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky034
更新日期:2018-04-06 00:00:00
abstract::FASTQ has emerged as a common file format for sharing sequencing read data combining both the sequence and an associated per base quality score, despite lacking any formal definition to date, and existing in at least three incompatible variants. This article defines the FASTQ format, covering the original Sanger stand...
journal_title:Nucleic acids research
pub_type: 历史文章,杂志文章,评审
doi:10.1093/nar/gkp1137
更新日期:2010-04-01 00:00:00
abstract::A physical map is presented for the 1200 kb genome of Mycoplasma mycoides subsp. mycoides Y, locating 32 cleavage sites for 8 restriction endonucleases. The large restriction fragments involved were separated and sized by pulsed-field agarose gel electrophoresis. Their locations on the map were determined by probing S...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/16.13.6027
更新日期:1988-07-11 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::Much effort has long been devoted to unraveling the coordinated cellular response to genotoxic insults. In view of the difficulty of obtaining human biological samples of homogeneous origin, I have established a set of stable human clones where one DNA repair gene has been stably silenced by means of RNA interference....
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkm195
更新日期:2007-01-01 00:00:00
abstract::The widespread applications of various 'omics' technologies in biomedical research together with the emergence of public data repositories have resulted in a plethora of data sets for almost any given physiological state or disease condition. Properly combining or integrating these data sets with similar basic hypothe...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt338
更新日期:2013-07-01 00:00:00
abstract::Phenotype analysis is commonly recognized to be of great importance for gaining insight into genetic interaction underlying inherited diseases. However, few computational contributions have been proposed for this purpose, mainly owing to lack of controlled clinical information easily accessible and structured for comp...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gki454
更新日期:2005-07-01 00:00:00
abstract::The determination of distances between specific points in nucleic acids is essential to understanding their behaviour at the molecular level. The ability to measure distances of 2-10 nm is particularly important: deformations arising from protein binding commonly fall within this range, but the reliable measurement of...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa086
更新日期:2020-04-06 00:00:00
abstract::Secondary structure-sensitive chemical and enzymatic probes have been used to produce a model for the folding of the last 68 residues of the 3'-non-coding region of beet necrotic yellow vein benevirus RNA 3. The structure consists of two stem-loops separated by a single-stranded region. RNA 3-derived transcripts were ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/25.23.4723
更新日期:1997-12-01 00:00:00
abstract::All exchangeable protons in a short DNA helix, d(CG)3 sodium salt, have been studied by proton nuclear magnetic resonance. The cytidine and guanosine amino protons have been assigned for the first time. As a function of temperature the cytidine amino protons and the imino protons behave very similarly, their relaxatio...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/12.21.8269
更新日期:1984-11-12 00:00:00
abstract::Spectroscopic studies of the complexes of double-stranded (ds) DNA with the polymethylene-amine linked heterodimers thiazole orange-thiazole blue, thiazole orange-ethidium, and fluorescein-ethidium, in each case show efficient energy transfer from donor to acceptor chromophores (Benson, S.C., Singh, P. and Glazer, A.N...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/21.24.5720
更新日期:1993-12-11 00:00:00
abstract::The U1A protein is a sequence-specific RNA binding protein found in the U1 snRNP particle where it binds to stem/loop II of U1 snRNA. U1A contains two 'RNP' or 'RRM' (RNA Recognition Motif) domains, which are common to many RNA-binding proteins. The N-terminal RRM has been shown to bind specifically to the U1 RNA stem...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/20.16.4283
更新日期:1992-08-25 00:00:00
abstract::The laboratory mouse is the premier animal model for studying human biology because all life stages can be accessed experimentally, a completely sequenced reference genome is publicly available and there exists a myriad of genomic tools for comparative and experimental research. In the current era of genome scale, dat...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gks1115
更新日期:2013-01-01 00:00:00
abstract::We have studied the role of the C-terminal domain of the alpha subunit (alphaCTD) of Escherichia coli RNA polymerase during transcription initiation at promoters lacking an UP-element. The temperature requirement for open complex formation was used as an indication of the kinetics of this process. We have previously s...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/27.9.2051
更新日期:1999-05-01 00:00:00
abstract::Cloned DNA from the EcoRI J fragment of EBV has been used as template for in vitro transcription experiments using cell-free extracts prepared from HeLa or KB cells. Two EBV specific RNAs each about 175 bases in length were synthesised and nuclease 51 mapping experiments determined that these in vitro products corresp...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/10.11.3407
更新日期:1982-06-11 00:00:00
abstract::Histone H2B O-GlcNAcylation is an important post-translational modification of chromatin during gene transcription. However, how this epigenetic modification is regulated remains unclear. Here we found that the energy-sensing adenosine-monophosphate-activated protein kinase (AMPK) could suppress histone H2B O-GlcNAcyl...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gku236
更新日期:2014-05-01 00:00:00
abstract::In vitro selection of RNA-cleaving DNAzymes is a powerful method for isolating metal-specific DNA. A few successful examples are known, but it is still difficult to target some thiophilic metals such as Cd(2+) due to limited functional groups in DNA. While using modified bases expands the chemical functionality of DNA...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv519
更新日期:2015-07-13 00:00:00
abstract::The gene-external part of the human 7S L promoter was analyzed by transcription in vitro and in vivo. Compared to the wild type promoter (-178), a -66 5'deletion mutant revealed full activity in vitro but was inefficiently transcribed in vivo. Further deletion to -37 reduced template activity to 50% in vitro and to ba...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.23.6779
更新日期:1990-12-11 00:00:00
abstract::A filter binding assay was developed that allows measurement of specific binding of trp repressor to operator DNA. The most important feature of this procedure is the concentration and type of salt present in the binding buffer. Using this assay the dissociation constant of the repressor-operator complex was determine...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/15.13.5339
更新日期:1987-07-10 00:00:00