Abstract:
:We describe a new exonuclease-based method for joining and/or constructing two or more DNA molecules. DNA fragments containing ends complementary to those of a vector or another independent molecules were generated by the polymerase chain reaction. The 3' ends of these molecules as well as the vector DNA were then recessed by exonuclease activity and annealed in an orientation-determined manner via their complementary single-stranded regions. This recombinant DNA can be transformed directly into bacteria without a further ligase-dependent reaction. Using this approach, we have constructed recombinant DNA molecules rapidly, efficiently and directionally. This method can effectively replace conventional protocols for PCR cloning, PCR SOEing, DNA subcloning and site-directed mutagenesis.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Yang YS,Watson WJ,Tucker PW,Capra JDdoi
10.1093/nar/21.8.1889subject
Has Abstractpub_date
1993-04-25 00:00:00pages
1889-93issue
8eissn
0305-1048issn
1362-4962journal_volume
21pub_type
杂志文章abstract::Natural antisense transcripts are at least partially complementary to their sense transcripts. Cis-Sense/Antisense pairs (cis-SAs) have been extensively characterized and known to play diverse regulatory roles, whereas trans-Sense/Antisense pairs (trans-SAs) in animals are poorly studied. We identified long trans-SAs ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkn470
更新日期:2008-09-01 00:00:00
abstract::The E. coli single-stranded binding protein (SSB) has been demonstrated in vitro to be involved in a number of replicative, DNA renaturation, and protective functions. It was shown previously that SSB can interact with exonuclease I to stimulate the hydrolysis of single-stranded DNA. We demonstrate here that E. coli S...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/22.2.247
更新日期:1994-01-25 00:00:00
abstract::The molecular organization of the dunce gene of Drosophila melanogaster has proved to be particularly complex, with two divergently transcribed genes, Sgs-4 and Pig-1, nested within its 79 kb intron (1). Here we report the identification and the molecular characterization of a third gene nested within the transcriptio...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.19.5837
更新日期:1990-10-11 00:00:00
abstract::Present results provide direct evidence of the nature of a conformational change in DNA when nucleosomes are formed from core histones and poly [d(A-T)]. First, we have found some features which have characteristic aspects of the A like conformation of DNA. Thus, an increased contribution due to a sugar conformation c...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/9.19.4879
更新日期:1981-10-10 00:00:00
abstract::A normal human population has been screened for the existence of further restriction fragment length polymorphisms (RFLPs) in the clotting factor IX gene in addition to the TaqI polymorphism already characterised (1,2). Two polymorphic loci were found, both within 6 Kb of the TaqI polymorphism within the body of the f...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/12.23.8861
更新日期:1984-12-11 00:00:00
abstract::Gene expression in Plasmodium falciparum is tightly regulated to ensure successful propagation of the parasite throughout its complex life cycle. The earliest transcriptomics studies in P. falciparum suggested a cascade of transcriptional activity over the course of the 48-hour intraerythrocytic developmental cycle (I...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx464
更新日期:2017-07-27 00:00:00
abstract::The Retinol-Binding Protein (RBP) is expressed primarily in the liver. The regulatory elements involved in its tissue-specific expression have been identified and mapped to the 5' flanking region of the RBP gene. In this paper heterokaryons and somatic cell-hybrids have been produced and analysed in order to demonstra...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.24.7235
更新日期:1990-12-25 00:00:00
abstract::The 'POU' (acronym of Pit-1, Oct-1, Unc-86) family of transcription factors share a common DNA-binding domain of approximately 160 residues, comprising so-called 'POUs' and 'POUh' sub-domains connected by a flexible linker. The importance of POU proteins as developmental regulators and tumor-promoting agents is due to...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkm453
更新日期:2007-01-01 00:00:00
abstract::We show that nucleic acid structures may be conveniently and inexpensively characterized by their UV thermal difference spectra. A thermal difference spectrum (TDS) is obtained for a nucleic acid by simply recording the ultraviolet absorbance spectra of the unfolded and folded states at temperatures above and below it...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gni134
更新日期:2005-09-12 00:00:00
abstract::Advances in single-cell RNA-sequencing techniques reveal the existence of distinct cell subpopulations. Identification of transcription factors (TFs) that define the identity of these subpopulations poses a challenge. Here, we postulate that identity depends on background subpopulations, and is determined by a synergi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkz147
更新日期:2019-04-23 00:00:00
abstract::The primary rRNA transcript contains a large external transcribed spacer (ETS) approximately 4,000 nucleotides in length. We have used subcloned DNA probes derived from the 5' end of the ETS in conjunction with Northern blot analysis of murine nuclear RNA to examine processing of this region. In agreement with the res...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/12.18.7187
更新日期:1984-09-25 00:00:00
abstract::The shape of the chromosomal DNA of the sulfur-dependent archaebacterium Sulfolobus acidocaldarius was analyzed by the pulsed-field gel electrophoresis(PFGE). S.acidocaldarius DNA digested with Notl showed two DNA bands at around 1.0 Mbp and 2.1 Mbp. Notl-linking clones were isolated from the library of S.acidocaldari...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.5.1133
更新日期:1990-03-11 00:00:00
abstract::We report here the construction of engineered endonuclease database (EENdb) (http://eendb.zfgenetics.org/), a searchable database and knowledge base for customizable engineered endonucleases (EENs), including zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs). EENs are artificial...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gks1144
更新日期:2013-01-01 00:00:00
abstract::Biological and chemical DNA fragmentation generates DNA molecules with a variety of termini, including blunt ends and single-stranded overhangs. We have developed a Next Generation Sequencing (NGS) assay, XACTLY, to interrogate the termini of fragmented DNA, information traditionally lost in standard NGS library prepa...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa128
更新日期:2020-05-07 00:00:00
abstract::Besides orthologs of other eukaryotic mismatch-repair (MMR) proteins, plants encode MSH7, a paralog of MSH6. The Arabidopsis thaliana recognition heterodimers AtMSH2*MSH6 (AtMutSalpha) and AtMSH2*MSH3 (AtMutSbeta) were previously found to bind the same subsets of mismatches as their counterparts in other eukaryotes--r...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkg780
更新日期:2003-10-15 00:00:00
abstract::Heat shock genes are activated by the binding of the heat shock transcription factor (HSF) to heat shock elements (HSEs), consisting of arrays of the 5-bp unit NGAAN arranged as inverted repeats. Here, we have investigated the interaction of the 5-bp unit with HSFs of Drosophila and Saccharomyces. Mutations within the...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/22.2.167
更新日期:1994-01-25 00:00:00
abstract::Highly conserved sequences present at an identical position near the 3' ends of eukaryotic and prokaryotic 5S rRNAs are complementary to the 5' strand of the m2(6)A hairpin structure near the 3' ends of 18S rRNA and 16S rRNA, respectively. The extent of base-pairing and the calculated stabilities of the hybrids that c...
journal_title:Nucleic acids research
pub_type: 杂志文章,评审
doi:10.1093/nar/7.7.1913
更新日期:1979-12-11 00:00:00
abstract::Current approaches to design efficient antisense RNAs (asRNAs) rely primarily on a thermodynamic understanding of RNA-RNA interactions. However, these approaches depend on structure predictions and have limited accuracy, arguably due to overlooking important cellular environment factors. In this work, we develop a bio...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx115
更新日期:2017-05-19 00:00:00
abstract::The EMBL Data Library was the first internationally supported central resource for nucleic acid sequence data. Working in close collaboration with its American counterpart, GenBank (1), the library prepares and makes available to the scientific community a comprehensive collection of the published nucleic acid sequenc...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/14.1.5
更新日期:1986-01-10 00:00:00
abstract::Herpes simplex virus-1 US11 is a RNA-binding protein with a novel RNA-binding domain. US11 has been reported to exhibit sequence- and conformation-specific RNA-binding, but the sequences and conformations important for binding are not known. US11 has also been described as a double-stranded RNA (dsRNA)-binding protein...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gki919
更新日期:2005-10-24 00:00:00
abstract::We took advantage of the unusual genomic organization of the ciliate Oxytricha trifallax to screen for eukaryotic non-coding RNA (ncRNA) genes. Ciliates have two types of nuclei: a germ line micronucleus that is usually transcriptionally inactive, and a somatic macronucleus that contains a reduced, fragmented and rear...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr501
更新日期:2011-09-01 00:00:00
abstract::The transmission of genetic information relies on Watson-Crick base pairing between nucleoside phosphates and template bases in template-primer complexes. Enzyme-free primer extension is the purest form of the transmission process, without any chaperon-like effect of polymerases. This simple form of copying of sequenc...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gku314
更新日期:2014-06-01 00:00:00
abstract::Previous studies of the epithelial specificity of the human papillomavirus type 16 (HPV-16) enhancer pointed out an important role of nuclear factor I (NFI). In epithelial cells, NFI proteins are derived from the NFI-C gene and referred to as NFI/CTF. In contrast, fibroblasts, where the enhancer is inactive, express h...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/22.19.3825
更新日期:1994-09-25 00:00:00
abstract::(19)F NMR spectroscopy has proved to be a valuable tool to monitor functionally important conformational transitions of nucleic acids. Here, we present a systematic investigation on the application of 5-fluoro pyrimidines to probe DNA and RNA secondary structures. Oligonucleotides with the propensity to adapt secondar...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkp862
更新日期:2009-12-01 00:00:00
abstract::Transcription and mRNA export are linked processes. However, the molecular mechanisms of this coordination are not clear. Sus1 (hENY2) participates in this coordination as part of two protein complexes: SAGA, a transcriptional co-activator; TREX-2, which functions in mRNA biogenesis and export. Here, we investigate th...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt272
更新日期:2013-06-01 00:00:00
abstract::Aberrant CpG methylation changes occurring during tumour progression include the loss (hypomethylation) and gain (hypermethylation) of methyl groups. Techniques currently available for examining such changes either require selection of a region, then examination of methylation changes, or utilise methylation-sensitive...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/29.24.e123
更新日期:2001-12-15 00:00:00
abstract::Mammalian antibody switch regions (∼1500 bp) are composed of a series of closely neighboring G4-capable sequences. Whereas numerous structural and genome-wide analyses of roles for minimal G4s in transcriptional regulation have been reported, Long G4-capable regions (LG4s)-like those at antibody switch regions-remain ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa357
更新日期:2020-06-19 00:00:00
abstract::We have used a polymerase chain reaction (PCR) procedure to analyse low abundance complementary sense RNAs of Digitaria streak virus (DSV) from infected leaves of Digitaria setigera. This study has confirmed that both spliced and unspliced RNAs are synthesised by the same transcription unit. The position of the intron...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.24.7259
更新日期:1990-12-25 00:00:00
abstract::HORMA domain-containing proteins (HORMADs) play an essential role in meiosis in many organisms. The meiotic HORMADs, including yeast Hop1, mouse HORMAD1 and HORMAD2, and Arabidopsis ASY1, assemble along chromosomes at early prophase and the closure motif at their C-termini has been hypothesized to be instrumental for ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa527
更新日期:2020-11-18 00:00:00
abstract::Deciphering gene regulatory network architecture amounts to the identification of the regulators, conditions in which they act, genes they regulate, cis-acting motifs they bind, expression profiles they dictate and more complex relationships between alternative regulatory partnerships and alternative regulatory motifs...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gki166
更新日期:2005-01-31 00:00:00