Abstract:
:We have developed efficient methods for creating artificial transposons and inserting these transposons into plasmid targets in vitro, primarily for the purpose of DNA mapping and sequencing. A novel plasmid has been engineered to convert virtually any DNA sequence, or combination of sequences, into an artificial transposon; hence, custom transposons containing any desired feature can be easily designed and constructed. Such transposons are then efficiently inserted into plasmid targets, in vitro, using the integrase activity present in yeast Ty1 virus-like particles. A single in vitro integration reaction, which resembles a simple restriction digestion in the complexity of the reaction, gives rise to thousands of recoverable insertion events within DNA target molecules; this frequency approaches one insertion per phosphodiester bond in typical plasmids. Importantly, transposon insertions are recovered from all regions of DNA inserts carried on plasmid targets, indicating that integration is a random or nearly-random process. Because of its versatility, this technology offers a generalized method of generating recombinant DNA molecules of a desired structure. We have adapted this system for DNA sequencing by developing a customized artificial transposon to insert new primer binding sites into internal regions of DNA inserts carried on cloning vectors. Transposon insertions have been generated throughout several different yeast and human DNA inserts carried on plasmids, allowing the efficient recovery of sequence information from these inserts. Our results demonstrate the overall utility of this method for both small and large-scale DNA sequencing, as well as general DNA restructuring, and indicate that it could be adapted for use with a number of additional applications including functional genetic analysis.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Devine SE,Boeke JDdoi
10.1093/nar/22.18.3765subject
Has Abstractpub_date
1994-09-11 00:00:00pages
3765-72issue
18eissn
0305-1048issn
1362-4962journal_volume
22pub_type
杂志文章abstract::In vivo transcripts of the L (4.5 kb) and M (1.9 kb) dsRNA plasmids were examined in type I killers of Saccharomyces cerevisiae. Transcripts for both plasmids include full-length (l,m) and partial-length (la,ma) single-stranded species. Both L-dsRNA transcripts (l,la) have in vitro mRNA activity for L-P1, previously s...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/11.4.1077
更新日期:1983-02-25 00:00:00
abstract::Transcription factors (TFs) are sequence-specific DNA-binding proteins acting as critical regulators of gene expression. The Plant Promoter Analysis Navigator (PlantPAN; http://PlantPAN2.itps.ncku.edu.tw) provides an informative resource for detecting transcription factor binding sites (TFBSs), corresponding TFs, and ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv1035
更新日期:2016-01-04 00:00:00
abstract::SynergyFinder (https://synergyfinder.fimm.fi) is a stand-alone web-application for interactive analysis and visualization of drug combination screening data. Since its first release in 2017, SynergyFinder has become a widely used web-tool both for the discovery of novel synergistic drug combinations in pre-clinical mo...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa216
更新日期:2020-07-02 00:00:00
abstract::The DNA photosensitisers m-iodo Hoechst and m-iodo, p-methoxy Hoechst have been co-crystallised with the oligonucleotide d(CGCGAATTCGCG)(2)and their crystal structures determined. The crystals were then subjected to slow dehydration, which reduced their solvent contents from 40 (normal) to 30 (partially dehydrated) an...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/28.5.1252
更新日期:2000-03-01 00:00:00
abstract::Recently, a family of transcription factors structurally related to Sp1 has been described; thus, more than one activator may bind to the GC boxes present in a number of viral and cellular promoters. We have compared the transactivation potentials of Sp1, Sp3 and Sp4 proteins on the human immunodeficiency virus type 1...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/22.23.4914
更新日期:1994-11-25 00:00:00
abstract::A truly universal nucleobase enables a host of novel applications such as simplified templates for PCR primers, randomized sequencing and DNA based devices. A universal base must pair indiscriminately to each of the canonical bases with little or preferably no destabilization of the overall duplex. In reality, many ca...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw736
更新日期:2016-10-14 00:00:00
abstract::The molecular mechanisms that underlie the neurological manifestations of patients with inherited diseases of vitamin B12 (cobalamin) metabolism remain to date obscure. We observed transcriptomic changes of genes involved in RNA metabolism and endoplasmic reticulum stress in a neuronal cell model with impaired cobalam...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky634
更新日期:2018-09-06 00:00:00
abstract::2'-Deoxyoligonucleotides with 5-fluorocytosine residues incorporated at specific positions of the nucleotide sequence are tools of great potential in the study of the catalytic mechanism by which DNA cytosine methyltransferases methylate the 5-position of DNA cytosine residues in specific sequence contexts. Chemical s...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/20.10.2421
更新日期:1992-05-25 00:00:00
abstract::Characterization of the ability of human interferons (IFNs) to rapidly induce genes led to the identification of the first two members of the STAT (signal transducers and activators of transcription) family, Stat1 and Stat2. To study the unique role of this transcription factor in IFN signaling under more physiologica...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/27.21.4191
更新日期:1999-11-01 00:00:00
abstract::The human 7SL RNA component of the signal recognition particle can be separated into four major conformers by nondenaturing polyacrylamide gel electrophoresis. We have investigated what sequences give 7SL RNA the property to exist in different conformations. The human 7SL gene 7L30.1 was mutagenized using the random l...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/14.11.4639
更新日期:1986-06-11 00:00:00
abstract::Terminal deoxynucleotidyl transferase, which requires a single-stranded DNA primer under the usual assay conditions, can be made to accept double-stranded DNA as primer for the addition of either rNMP or dNMP, if Mg+2 ion is replaced by Co+2 ion. The priming efficiency in the presence of (C leads to) CO+2 ion with res...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/3.1.101
更新日期:1976-01-01 00:00:00
abstract::We studied in vivo transcription and gene expression directed by a series of synthetic sequences, bearing the consensus hexamer (CH) pair of E.coli promoters in various contexts. The results demonstrate that, for the contexts tested, the CH pair supports transcription activity and gene expression, whether the spacer l...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.5.1137
更新日期:1990-03-11 00:00:00
abstract::The APOBEC3 proteins are unique to mammals. Many inhibit retrovirus infection through a cDNA cytosine deamination mechanism. HIV-1 neutralizes this host defense through Vif, which triggers APOBEC3 ubiquitination and degradation. Here, we report an APOBEC3F-like, double deaminase domain protein from three artiodactyls:...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkl721
更新日期:2006-01-01 00:00:00
abstract::Large expansions of the trinucleotide repeat GAA*TTC within the first intron of the X25 (frataxin) gene cause Friedreich's ataxia, the most common inherited ataxia. Expansion leads to reduced levels of frataxin mRNA in affected individuals. Here we show that GAA*TTC tracts, in the absence of any other frataxin gene se...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/28.14.2815
更新日期:2000-07-15 00:00:00
abstract::Two new crescent-shaped unsymmetrical cyanine dyes have been synthesised and their interactions with DNA have been investigated by different spectroscopic methods. These dyes are analogues to a minor groove binding unsymmetrical cyanine dye, BEBO, recently reported by us. In this dye, the structure of the known interc...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkg821
更新日期:2003-11-01 00:00:00
abstract::Melanotransferrin (MTf) is a tumor associated antigen found in abundance on the surface of melanoma cells. It is a transferrin-like protein found in low amount in most adult tissues and whose gene is reminiscent of house-keeping genes. With the goal of understanding the regulatory mechanisms which might explain the en...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/20.11.2853
更新日期:1992-06-11 00:00:00
abstract::We describe the unique features of an aberrantly rearranged mu immunoglobulin heavy chain gene isolated from MPC-11 cells (a gamma 2b producing Balb/c plasmacytoma). A novel rearrangement has occurred 1.5 Kb 5' of the MPC-11 mu gene (denoted 18b mu) resulting in the deletion of the majority of the repetitive switch re...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/10.23.7751
更新日期:1982-12-11 00:00:00
abstract::Human stem cells are promising sources for regenerative therapy. To ensure safety of future therapeutic applications, the differentiation potency of stem cells has to be tested and be widely opened to the public. The potency is generally assessed by teratoma formation comprising differentiated cells from all three ger...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv1096
更新日期:2016-01-04 00:00:00
abstract::Synthesis and antisense activity of oligonucleotides modified with 2'-O-[2-[(N,N-dimethylamino)oxy] ethyl] (2'-O-DMAOE) are described. The 2'-O-DMAOE-modified oligonucleotides showed superior metabolic stability in mice. The phosphorothioate oligonucleotide 'gapmers', with 2'-O-DMAOE- modified nucleoside residues at t...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh220
更新日期:2004-02-03 00:00:00
abstract::Since July 1995, the European Bioinformatics Institute (EBI) has maintained RHdb (http://www.ebi.ac.uk/RHdb/RHdb.html ), a public database for radiation hybrid data. Radiation hybrid mapping is an important technique for determining high resolution maps. Recently, CORBA access has been added to Rhdb. The EBI is an Out...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.1.102
更新日期:1998-01-01 00:00:00
abstract::A cDNA that encodes a transcription factor able to recognize the thyroglobulin gene promoter in vitro was isolated from a dog thyroid cDNA expression library in lambda gt11. The library was screened with a multimerized 20 bp-oligonucleotide probe corresponding to the -126 to -107 bp region of the bovine thyroglobulin ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/19.5.1121
更新日期:1991-03-11 00:00:00
abstract::A minor tRNA-Glu1 constituent of a preparation of E. coli B tRNA-Glu (Oak Ridge) has a guanosine residue at position 66 rather than an adenosine as in the tRNA-Glu2 described by Ohashi et al. (3). Automated sequential degradation was used to sequence this region. ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/2.4.469
更新日期:1975-04-01 00:00:00
abstract::A 2.1-kb cDNA clone representing a new member of the protease inhibitor family was isolated from a human liver cDNA library. The inhibitor, named human Leuserpin 2 (hLS2), comprises 480 amino acids and contains a leucine residue at its putative reactive center. HLS2 is about 25-28% homologous to three human members of...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/14.2.1073
更新日期:1986-01-24 00:00:00
abstract::Here we report that the poor binding of methylphosphonate oligodeoxynucleosides (MP-ODNs) to their nucleic acid targets can be improved by additional inversion of the anomeric configuration (from beta to alpha) in the sugar moieties to give a new class of analogs, MP alpha-oligonucleosides. MP alpha-dT12and MP 5' alph...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.20.4551
更新日期:1998-10-15 00:00:00
abstract::Adaptation to the host cell environment to efficiently take-over the host cell's machinery is crucial in particular for small RNA viruses like picornaviruses that come with only small RNA genomes and replicate exclusively in the cytosol. Their Internal Ribosome Entry Site (IRES) elements are specific RNA structures th...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gks182
更新日期:2012-07-01 00:00:00
abstract::Recent effective use of TAL Effectors (TALEs) has provided an important approach to the design and synthesis of sequence-specific DNA-binding proteins. However, it is still a challenging task to design and manufacture effective TALE modulators because of the limited knowledge of TALE-DNA interactions. Here we synthesi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gku535
更新日期:2014-08-01 00:00:00
abstract::The synthesis of cyclic di-, tri-, tetra-, penta- and hexa-thymidylic acids (16; n = 2, 3, 4, 5 and 6) and the cyclic hexadeoxyribonucleotide [d(CpCpTpApGpGp)], via the "filtration" approach, is reported. Some of the physical properties of the cyclic oligonucleotides are discussed, and their susceptibility to digestio...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/17.20.8221
更新日期:1989-10-25 00:00:00
abstract::chipD is a web server that facilitates design of DNA oligonucleotide probes for high-density tiling arrays, which can be used in a number of genomic applications such as ChIP-chip or gene-expression profiling. The server implements a probe selection algorithm that takes as an input, in addition to the target sequences...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq517
更新日期:2010-07-01 00:00:00
abstract::We previously proposed that the first enzyme for histidine biosynthesis in Salmonellatyphimurium plays a role in regulating expression of the histidine operon and that in order to play this role the enzyme must form a complex with histidyl-tRNA. Among the many observations that led to these conclusions were 1) that re...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/1.7.881
更新日期:1974-07-01 00:00:00
abstract::Most mutations in cancer genomes occur in the non-coding regions with unknown impact on tumor development. Although the increase in the number of cancer whole-genome sequences has revealed numerous putative non-coding cancer drivers, their information is dispersed across multiple studies making it difficult to underst...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa915
更新日期:2021-01-08 00:00:00