Abstract:
:We have adapted a method for making libraries of mutations in any specific gene for use in the fission yeast Schizosaccharomyces pombe . This elegant and simple method consists of PCR amplification of the gene of interest, followed by co-transformation of fission yeast with the PCR fragment and a linearized plasmid vector prepared such that the ends of the vector share DNA sequence with the ends of the PCR fragment. Homologous recombination between the vector and the PCR fragment occurs at a high frequency and results in a collection of yeast transformants, most harboring a mutated allele of the original gene within the vector of choice. This library can then be screened or selected for phenotypes of interest.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Kostrub CF,Lei EP,Enoch Tdoi
10.1093/nar/26.20.4783subject
Has Abstractpub_date
1998-10-15 00:00:00pages
4783-4issue
20eissn
0305-1048issn
1362-4962pii
gkb741journal_volume
26pub_type
杂志文章abstract::Restarting stalled replication forks is vital to avoid fatal replication errors. Previously, it was demonstrated that hydroxyurea-stalled replication forks rescue replication either by an active restart mechanism or by new origin firing. To our surprise, using the DNA fibre assay, we only detect a slightly reduced for...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr420
更新日期:2011-09-01 00:00:00
abstract::Detection of cancer-associated somatic mutations has broad applications for oncology and precision medicine. However, this becomes challenging when cancer-derived DNA is in low abundance, such as in impure tissue specimens or in circulating cell-free DNA. Next-generation sequencing (NGS) is particularly prone to techn...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkz474
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abstract::The numerous genomic sequences and ESTs released by the Arabidopsis thaliana Genome Initiative (AGI) have allowed a systematic and functional study of the DEAD box RNA helicase family. Sequencing and in silico analysis led to the characterization of 28 novel A. thaliana DEAD box RNA helicases forming a family of 32 me...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/27.2.628
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abstract::A mass culture of human diploid fibroblasts, and eight clones isolated from that mass culture, were examined for methylation patterns in several regions of DNA. Plasmid-inserted cDNA sequences were used as probes for alpha-hCG, beta-globin, A gamma- and G gamma-globin, and beta- and gamma-actin gene regions. Each prob...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/10.14.4293
更新日期:1982-07-24 00:00:00
abstract::We define phosphovariants as genetic variations that change phosphorylation sites or their interacting kinases. Considering the essential role of phosphorylation in protein functions, it is highly likely that phosphovariants change protein functions. Therefore, a comparison of phosphovariants between individuals or be...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkn1008
更新日期:2009-03-01 00:00:00
abstract::For sophisticated gene targeting procedures requiring two sequential selective steps to operate efficiently it is essential that the marker genes used are not prone to position effects. The double replacement gene targeting procedure, to produce mice with subtle gene alterations, is based on the use of hypoxanthine ph...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/25.19.3937
更新日期:1997-10-01 00:00:00
abstract::Dnmt3a and Dnmt3b are paralogous enzymes responsible for de novo DNA methylation but with distinguished biological functions. In mice, disruption of Dnmt3b but not Dnmt3a causes global DNA hypomethylation, especially in repetitive sequences, which comprise the large majority of methylated DNA in the genome. By measuri...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq456
更新日期:2010-10-01 00:00:00
abstract::In this study, we present the DNA-Binding Site Identifier (DBSI), a new structure-based method for predicting protein interaction sites for DNA binding. DBSI was trained and validated on a data set of 263 proteins (TRAIN-263), tested on an independent set of protein-DNA complexes (TEST-206) and data sets of 29 unbound...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt617
更新日期:2013-09-01 00:00:00
abstract::To organize data resulting from the phenotypic characterization of a library of 30,000 T-DNA enhancer trap (ET) insertion lines of rice (Oryza sativa L cv. Nipponbare), we developed the Oryza Tag Line (OTL) database (http://urgi.versailles.inra.fr/OryzaTagLine/). OTL structure facilitates forward genetic search for sp...
journal_title:Nucleic acids research
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更新日期:2008-01-01 00:00:00
abstract::We describe an improved model of DNA replication in Xenopus egg extracts, in which a circular plasmid immobilized on paramagnetic beads is used as a template. DNA synthesis occurred on either circular or linear plasmids coupled to the beads, but only DNA synthesis on the circular plasmid was inhibited by geminin and a...
journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:2006-07-26 00:00:00
abstract::Guanine-rich DNA sequences tend to form four-stranded G-quadruplex structures. Characteristic glycosidic conformational patterns along the G-strands, such as the 5'-syn-anti-syn-anti pattern observed with the Oxytricha nova telomeric G-quadruplexes, have been well documented. However, an explanation for these featured...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr031
更新日期:2011-05-01 00:00:00
abstract::The high mobility group protein (HMG)-box is a DNA-binding domain found in many proteins that bind preferentially to DNA of irregular structures in a sequence-independent manner and can bend the DNA. We show here that GST-fusion proteins of HMG domains from HMG1 and HMG2 promote a triple-stranded complex formation bet...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/24.23.4733
更新日期:1996-12-01 00:00:00
abstract::In testis mRNA stability and translation initiation are extensively under the control of poly(A)-binding proteins (PABP). Here we have cloned a new human testis-specific PABP (PABP3) of 631 amino acids (70.1 kDa) with 92.5% identical residues to the ubiquitous PABP1. A northern blot of multiple human tissues hybridise...
journal_title:Nucleic acids research
pub_type: 杂志文章
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abstract::The oncogenic transformation of normal cells into malignant, rapidly proliferating cells requires major alterations in cell physiology. For example, the transformed cells remodel their metabolic processes to supply the additional demand for cellular building blocks. We have recently demonstrated essential metabolic pr...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv1337
更新日期:2016-01-04 00:00:00
abstract::Many multimeric transcription factors recognize DNA sequence patterns by cooperatively binding to bipartite elements composed of half sites separated by a flexible spacer. We developed a novel bipartite algorithm, bipartite pattern discovery (Bipad), which produces a mathematical model based on information maximizatio...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh825
更新日期:2004-09-23 00:00:00
abstract::Mutator-like transposable elements (MULEs) are widespread across fungal, plant and animal species. Despite their abundance and importance as genetic tools in plants, the transposition mechanism of the MULE superfamily was previously unknown. Discovery of the Muta1 element from Aedes aegypti and its successful transpos...
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更新日期:2017-06-20 00:00:00
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journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/21.25.5896
更新日期:1993-12-25 00:00:00
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journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv563
更新日期:2015-10-15 00:00:00
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
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abstract::A novel expression system was established that allows expression and propagation of the human hepatitis B virus (HBV) genome in cultured animal cells. An overlength HBV genome encoding the viral pregenomic RNA was put under transcriptional control of the human metallothionein IIA promoter thereby replacing the endogen...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/15.24.10117
更新日期:1987-12-23 00:00:00
abstract::The anti-cancer drug 2',2'-difluoro-2'-deoxycytidine (dFdC) is internally incorporated into DNA in vitro. To determine the effects of this incorporation on DNA structure and function, the beta-cyanoethyl phosphoramidite of dFdC was synthesized and oligodeoxyribonucleotides containing dFdC were made using automated sol...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/20.7.1763
更新日期:1992-04-11 00:00:00
abstract::Structural domains are considered as the basic units of protein folding, evolution, function and design. Automatic decomposition of protein structures into structural domains, though after many years of investigation, remains a challenging and unsolved problem. Manual inspection still plays a key role in domain decomp...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkg189
更新日期:2003-02-01 00:00:00
abstract::A requirement for specific RNA folding is that the free-energy landscape discriminate against non-native folds. While tertiary interactions are critical for stabilizing the native fold, they are relatively non-specific, suggesting additional mechanisms contribute to tertiary folding specificity. In this study, we use ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv1055
更新日期:2016-01-08 00:00:00
abstract::Synchronous plasmodia of Physarum polycephalum in DNA synthesis were pulse-labelled with [oH]- thymidine for time periods of 15 seconds up to 9 minutes, or given a 30 seconds pulse followed by chase periods of 9 minutes up to 6 hours. Sedimentation analysis in alkaline sucrose gradients revealed at least five species ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/2.8.1381
更新日期:1975-08-01 00:00:00
abstract::The rpoA gene, encoding the alpha subunit of RNA polymerase, was cloned from Streptomyces coelicolor A3(2). It is preceded by rpsK and followed by rplQ, encoding ribosomal proteins S11 and L17, respectively, similar to the gene order in Bacillus subtilis. The rpoA gene specifies a protein of 339 amino acids with deduc...
journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:1996-11-15 00:00:00
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journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/27.1.95
更新日期:1999-01-01 00:00:00
abstract::In prokaryotes and archaea transfer ribonucleic acid (tRNA) stability as well as cellular UV protection relies on the post-transcriptional modification of uracil at position 8 (U8) of tRNAs by the 4-thiouridine synthetase ThiI. Here, we report three crystal structures of ThiI from Thermotoga maritima in complex with a...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gku249
更新日期:2014-06-01 00:00:00