Abstract：
:A new procedure has been developed for the efficient cloning of complex PCR mixtures, resulting in libraries exclusively consisting of recombinant clones. Recombinants are generated between PCR products and a PCR-amplified plasmid vector. The procedure does not require the use of restriction enzymes, T4 DNA ligase or alkaline phosphatase. The 5'-ends of the primers used to generate the cloneable PCR fragments contain an additional 12 nucleotide (nt) sequence lacking dCMP. As a result, the amplification products include 12-nt sequences lacking dGMP at their 3'-ends. The 3'-terminal sequence can be removed by the action of the (3'----5') exonuclease activity of T4 DNA polymerase in the presence of dGTP, leading to fragments with 5'-extending single-stranded (ss) tails of a defined sequence and length. Similarly, the entire plasmid vector is amplified with primers homologous to sequences in the multiple cloning site. The vector oligos have additional 12-nt tails complementary to the tails used for fragment amplification, permitting the creation of ss-ends with T4 DNA polymerase in the presence of dCTP. Circularization can occur between vector molecules and PCR fragments as mediated by the 12-nt cohesive ends, but not in mixtures lacking insert fragments. The resulting circular recombinant molecules do not require in vitro ligation for efficient bacterial transformation. We have applied the procedure for the cloning of inter-ALU fragments from hybrid cell-lines and human cosmid clones.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Aslanidis C,de Jong PJdoi
10.1093/nar/18.20.6069subject
Has Abstractpub_date
1990-10-25 00:00:00pages
6069-74issue
20eissn
0305-1048issn
1362-4962journal_volume
18pub_type
杂志文章abstract：:Topoisomerase II is an essential enzyme that is required for virtually every process that requires movement of DNA within the nucleus or the opening of the double helix. This enzyme helps to regulate DNA under- and overwinding and removes knots and tangles from the genetic material. In order to carry out its critical ...
journal_title：Nucleic acids research
pub_type： 杂志文章,评审
doi：10.1093/nar/gkn937
更新日期：2009-02-01 00:00:00
abstract：:Invariance of temporal order of genome replication in eukaryotic cells and its correlation with gene activity has been well-documented. However, recent data suggest a relax control of replication timing. To evaluate replication schedule accuracy, we detailed the replicational organization of the developmentally regula...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkm586
更新日期：2007-01-01 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：:Brome mosaic virus (BMV) is a tripartite genome, positive-sense RNA virus of plants. Previously it was demonstrated that local hybridization between BMV RNAs (RNA-RNA heteroduplex formation) efficiently promotes non-homologous RNA recombination. In addition, studies on the role of the BMV polymerase in RNA recombinati...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/28.8.1714
更新日期：2000-04-15 00:00:00
abstract：:Long Interspersed Element-1 (LINE-1) retrotransposition contributes to inter- and intra-individual genetic variation and occasionally can lead to human genetic disorders. Various strategies have been developed to identify human-specific LINE-1 (L1Hs) insertions from short-read whole genome sequencing (WGS) data; howev...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkz1173
更新日期：2020-02-20 00:00:00
abstract：:An 11 S ribonucleoprotein particle was isolated from maize endosperm and shown to be functionally and structurally equivalent to the mammalian signal recognition particle. However, unlike animal cells which apparently contain a single 7 SL RNA species, maize endosperm contains a heterogeneous population of 7 SL RNA. T...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/17.4.1573
更新日期：1989-02-25 00:00:00
abstract：:Topoisomerases are ubiquitous proteins found in all three domains of life. They change the topology of DNA via transient breaks on either one or two of the DNA strands to allow passage of another single or double DNA strand through the break. Topoisomerases are classified into two types: type I enzymes cleave one DNA ...
journal_title：Nucleic acids research
pub_type： 杂志文章,评审
doi：10.1093/nar/gkn1009
更新日期：2009-02-01 00:00:00
abstract：:The proposition that the 2-amino group of guanine plays a critical role in determining how antibiotics recognise their binding sites in DNA has been tested by relocating it, using tyrT DNA derivative molecules substituted with inosine plus 2,6-diaminopurine (DAP). Irrespective of their mode of interaction with DNA, su...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/23.6.885
更新日期：1995-03-25 00:00:00
abstract：:Bursts of free radicals produced by ionization of water in close vicinity to DNA can produce clusters of opposed DNA lesions and these are termed multiply damaged sites (MDS). How MDS are processed by the Escherichia coli DNA glycosylases, endonuclease (endo) III and endo VIII, which recognize oxidized pyrimidines, is...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/26.4.932
更新日期：1998-02-15 00:00:00
abstract：:UNITE (https://unite.ut.ee/) is a web-based database and sequence management environment for the molecular identification of fungi. It targets the formal fungal barcode-the nuclear ribosomal internal transcribed spacer (ITS) region-and offers all ∼1 000 000 public fungal ITS sequences for reference. These are clustere...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gky1022
更新日期：2019-01-08 00:00:00
abstract：:This study describes a novel functional property of the eukaryotic transcription factor NF-kappa B in altering the DNA structure. We observed that the binding of purified NF-kappa B to DNA was facilitated by spermine and cations which are known to promote DNA bending, and by a nick in one position at the 3'-end of the...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/18.22.6497
更新日期：1990-11-25 00:00:00
abstract：:The disappearance of defined restriction fragments of the beta 1-globin, an albumin and the A1 vitellogenin gene was quantitated after DNase I digestion and expressed by a sensitivity factor defined by a mathematical model. Analysis of naked DNA showed that the gene fragments have similar but not identical sensitivity...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/9.11.2455
更新日期：1981-06-11 00:00:00
abstract：:Hfq, a bacterial RNA-binding protein, was recently shown to contain the Sm1 motif, a characteristic of Sm and LSm proteins that function in RNA processing events in archaea and eukaryotes. In this report, comparative structural modeling was used to predict a three-dimensional structure of the Hfq core sequence. The pr...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkf508
更新日期：2002-09-01 00:00:00
abstract：:Long terminal repeat (LTR)-retrotransposons comprise a significant portion of the rice genome. Their complete characterization is thus necessary if the sequenced genome is to be annotated correctly. In addition, because LTR-retrotransposons can influence the expression of neighboring genes, the complete identification...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkl780
更新日期：2007-01-01 00:00:00
abstract：:We have isolated a pigment cell-specific cDNA clone from a B16 mouse melanoma cDNA library by differential hybridization. The mRNA of isolated cDNA is highly expressed in B16 melanoma cells and in black mouse (C57BL/6) skin, but is not detectable in mouse neuroblastoma cells nor in K1735 mouse amelanotic melanoma cell...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/14.6.2413
更新日期：1986-03-25 00:00:00
abstract：:In Pseudomonas aeruginosa the RNA chaperone Hfq and the catabolite repression control protein (Crc) act as post-transcriptional regulators during carbon catabolite repression (CCR). In this regard Crc is required for full-fledged Hfq-mediated translational repression of catabolic genes. RNAseq based transcriptome anal...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkx1245
更新日期：2018-02-16 00:00:00
abstract：:Stress response mechanisms that modulate the dynamics of tRNA degradation and accumulation from the cytoplasm to the nucleus have been studied in yeast, the rat hepatoma and human cells. In the current study, we investigated tRNA degradation and accumulation in HeLa cells under various forms of stress. We found that i...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkt153
更新日期：2013-04-01 00:00:00
abstract：:Starting from a crude E. coli extract, two activities which specifically protect highly repetitive bacterial DNA sequences (called PU for Palindromic Unit or REP for Repetitive Extragenic Palindromic sequence) against a digestion with Exonuclease III have been purified. We show that one of these activities is due to t...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/18.13.3941
更新日期：1990-07-11 00:00:00
abstract：:Proteins interacting with pre-mRNAs during early stages of spliceosome formation in a HeLa nuclear extract were investigated by photochemical RNA-protein crosslinking. The level of protein crosslinking to a beta-globin pre-mRNA was positively correlated with the presence of an intron. Proteins of 110,000, 59,000 and 3...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/18.11.3307
更新日期：1990-06-11 00:00:00
abstract：:Activating sequence factor 1 (ASF-1) is a nuclear DNA-binding activity that is found in monocots and dicots. It interacts with several TGACG-containing elements that have been characterized from viral and T-DNA genes, the prototypes of which are the as-1 element of the CaMV 35S promoter and the ocs element from the oc...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/23.18.3778
更新日期：1995-09-25 00:00:00
abstract：:The environmental carcinogen benzo[a]pyrene (BP) is metabolized to reactive diol epoxides that bind to cellular DNA by predominantly forming N2-guanine adducts (G*). Mutation hotspots for these adducts are frequently found in 5'- ... GG ... dinucleotide sequences, but their origins are poorly understood. Here we used ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkm022
更新日期：2007-01-01 00:00:00
abstract：:FlyBase is a database of genetic and molecular data concerning Drosophila. FlyBase is maintained as a relational database (in Sybase). The scope of FlyBase includes: genes, alleles (and phenotypes), aberrations, pointers to sequence data, clones, stock lists, Drosophila workers and bibliographic references. FlyBase is...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/24.1.53
更新日期：1996-01-01 00:00:00
abstract：:Regulation of gene transcription in both prokaryotes and eukaryotes involves formation of various DNA-multiprotein complexes of higher order structure through communication between distant regions of DNA. The communication between distant DNA sites occurs by interaction between proteins bound to the sites by looping o...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/25.4.873
更新日期：1997-02-15 00:00:00
abstract：:The macronuclear genome of Euplotes crassus contains two different genes, EFA1 and EFA2, encoding EF-1alpha proteins. They are derived from micronuclear precursors in the course of a sexual process termed conjugation. We have found that two apparent micronuclear precursors exist for EFA1. They differ in their potentia...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/24.16.3195
更新日期：1996-08-15 00:00:00
abstract：:SplicePort is a web-based tool for splice-site analysis that allows the user to make splice-site predictions for submitted sequences. In addition, the user can also browse the rich catalog of features that underlies these predictions, and which we have found capable of providing high classification accuracy on human s...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkm407
更新日期：2007-07-01 00:00:00
abstract：:Escherichia coli MutY is an adenine and a weak guanine DNA glycosylase involved in reducing mutagenic effects of 7,8-dihydro-8-oxo-guanine (8-oxoG). The C-terminal domain of MutY is required for 8-oxoG recognition and is critical for mutation avoidance of oxidative damage. To determine which residues of this domain ar...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkg434
更新日期：2003-06-15 00:00:00
abstract：:Chemically modified siRNAs are expected to have resistance toward nuclease degradation and good thermal stability in duplex formation for in vivo applications. We have recently found that 2'-OMe-4'-thioRNA, a hybrid chemical modification based on 2'-OMeRNA and 4'-thioRNA, has high hybridization affinity for complement...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gks204
更新日期：2012-07-01 00:00:00
abstract：:During erythropoiesis in the chick, the nucleosome repeat length increases from 190 base pairs to 212 base pairs. This increase is correlated with a dramatic increase in the concentration of the red cell specific histone H5 (from 0.2 molecules per nucleosome to 1 molecule per nucleosome) and with no change in the conc...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/5.4.1179
更新日期：1978-04-01 00:00:00
abstract：:Trinucleotide exchange (TriNEx) is a method for generating novel molecular diversity during directed evolution by random substitution of one contiguous trinucleotide sequence for another. Single trinucleotide sequences were deleted at random positions in a target gene using the engineered transposon MuDel that were su...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkn358
更新日期：2008-08-01 00:00:00
abstract：:Little is known about the function of most non-coding RNAs (ncRNAs). The majority of long ncRNAs (lncRNAs) is expressed at very low levels and it is a matter of intense debate whether these can be of functional relevance. Here, we identified lncRNAs regulating the viability of lung cancer cells in a high-throughput RN...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkx076
更新日期：2017-05-19 00:00:00
