Abstract：
:Although the thermophilic bacterium Thermus aquaticus grows optimally at 70 degrees C and cannot grow at moderate temperatures, its DNA polymerase I has significant activity at 20-37 degrees C. This activity is a bane to some PCRs, since it catalyzes non-specific priming. We report mutations of Klentaq (an N-terminal deletion variant) DNA polymerase that have markedly reduced activity at 37 degrees C yet retain apparently normal activity at 68 degrees C and resistance at 95 degrees C. The first four of these mutations are clustered on the outside surface of the enzyme, nowhere near the active site, but at the hinge point of a domain that has been proposed to move at each cycle of nucleotide incorporation. We show that the novel cold-sensitive mutants can provide a hot start for PCR and exhibit slightly improved fidelity.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Kermekchiev MB,Tzekov A,Barnes WMdoi
10.1093/nar/gkg813keywords:
subject
Has Abstractpub_date
2003-11-01 00:00:00pages
6139-47issue
21eissn
0305-1048issn
1362-4962journal_volume
31pub_type
杂志文章abstract：:Boundary elements partition eukaryotic chromatin into active and repressive domains, and can also block regulatory interactions between domains. Boundary elements act via diverse mechanisms making accurate feature-based computational predictions difficult. Therefore, we developed an unbiased algorithm that predicts th...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkr750
更新日期：2012-01-01 00:00:00
abstract：:X-linked agammaglobulinemia (XLA) is an immunodeficiency caused by mutations in the gene coding for Bruton's agammaglobulinemia tyrosine kinase (BTK). A database (BTKbase) of BTK mutations has been compiled and the recent update lists 368 entries from 318 unrelated families showing 228 unique molecular events. In addi...
journal_title：Nucleic acids research
pub_type： 杂志文章,多中心研究
doi：10.1093/nar/25.1.166
更新日期：1997-01-01 00:00:00
abstract：:Two new beta-lactoglobulin (BLG)/human serum albumin (HSA) hybrid gene vectors were constructed and tested for expression in COS-7 cells and in transgenic mice. The HSA sequences were inserted between the second and sixth BLG exons. Transient transfection experiments with these vectors as well as a series of additiona...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/24.4.602
更新日期：1996-02-15 00:00:00
abstract：:Molecular mechanisms that regulate gene expression can occur either before or after transcription. The information for post-transcriptional regulation can lie within the sequence or structure of the RNA transcript and it has been proposed that G-quadruplex nucleic acid sequence motifs may regulate translation as well ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkn511
更新日期：2008-11-01 00:00:00
abstract：:Metagenomics is a relatively recently established but rapidly expanding field that uses high-throughput next-generation sequencing technologies to characterize the microbial communities inhabiting different ecosystems (including oceans, lakes, soil, tundra, plants and body sites). Metagenomics brings with it a number ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkt961
更新日期：2014-01-01 00:00:00
abstract：:The major surface antigen of procyclic and epimastigote forms of Trypanosoma congolense in the tsetse fly is GARP (glutamic acid/alanine-rich protein), which is thought to be the analogue of procyclin/PARP in Trypanosoma brucei. We have studied two T.congolense GARP loci (the 4.3 and 4.4 loci) whose transcription is a...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/24.2.272
更新日期：1996-01-15 00:00:00
abstract：:The UCSC Genome Browser (https://genome.ucsc.edu) provides a web interface for exploring annotated genome assemblies. The assemblies and annotation tracks are updated on an ongoing basis-12 assemblies and more than 28 tracks were added in the past year. Two recent additions are a display of CRISPR/Cas9 guide sequences...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkx1020
更新日期：2018-01-04 00:00:00
abstract：:To elucidate the response to oxidative stress in eukaryotic cells, the effect of an oxidized nucleotide, 8-oxo-2'-deoxyguanosine 5'-triphosphate (8-oxo-dGTP), generated from dGTP with an active oxygen, on DNA synthesis was studied using a cell-free DNA replication system derived from Xenopus egg lysates with a single-...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/30.2.569
更新日期：2002-01-15 00:00:00
abstract：:Short DNA sequence motifs have been identified in viral and cellular enhancers which represent the binding sites for a variety of trans- acting factors. One such HeLa cell factor, EBP1, has been purified and shown to bind to sequences in the SV40 enhancer. The PRDII element in the human beta-interferon gene regulatory...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/17.2.499
更新日期：1989-01-25 00:00:00
abstract：:Based on parameters governing promoter activity and using regulatory elements of the lac, ara and tet operon transcription control sequences were composed which permit the regulation in Escherichia coli of several gene activities independently and quantitatively. The novel promoter PLtetO-1 allows the regulation of ge...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/25.6.1203
更新日期：1997-03-15 00:00:00
abstract：:The possibility of cloning large segments of DNA in cosmid vectors offers distinct advantages, in particular for the study of multigene families. Large size fragments of mouse embryo DNA were successfully cloned in the cosmid pHC 79. Twelve recombinants hybridizing specifically to an immunoglobulin kappa chain variabl...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/9.12.2777
更新日期：1981-06-25 00:00:00
abstract：:The detailed principles of the hierarchical folding of eukaryotic chromosomes have been revealed during the last two decades. Along with structures composing three-dimensional (3D) genome organization (chromatin compartments, topologically associating domains, chromatin loops, etc.), the molecular mechanisms that are ...
journal_title：Nucleic acids research
pub_type： 杂志文章,评审
doi：10.1093/nar/gkaa261
更新日期：2020-05-21 00:00:00
abstract：:The covalent attachment of thiol-modified DNA oligomers; to self-assembled monolayer silane films on fused silica and oxidized silicon substrates is described. A heterobifunctional crosslinking molecule bearing both thiol- and amino-reactive moieties was used to tether a DNA oligomer (modified at its terminus with a t...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/24.15.3031
更新日期：1996-08-01 00:00:00
abstract：:In a pilot study on SAGE on Reed-Sternberg cells we have sequenced 1055 tags representing 701 genes. Screening of the GenBank database resulted in the identification of a corresponding gene or EST for 490 of them. For 211 of the tags no homology could be detected. A major problem of the serial analysis of gene express...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/27.17.e17
更新日期：1999-09-01 00:00:00
abstract：:We developed a sensitive 32P-postlabeling method for the detection of bifunctional intrastrand crosslinks d(Pt-GpG) and d(Pt-ApG) in DNA in vitro and in vivo. After enzymatic digestion of DNA the positively charged platinum adducts were purified from unplatinated products, using strong cation exchange chromatography. ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/23.8.1300
更新日期：1995-04-25 00:00:00
abstract：:Aberrant DNA methylation at CpG dinucleotides can result in epigenetic silencing of tumour suppressor genes and represents one of the earliest events in tumourigenesis. To date, however, high-throughput tools that are capable of surveying the methylation status of multiple gene promoters have been restricted to a limi...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gni072
更新日期：2005-04-28 00:00:00
abstract：:RNA is ideally suited for in vitro evolution experiments, because a single RNA molecule possesses both genotypic (replicable sequence) and phenotypic (selectable shape) properties. Using systematic evolution of ligands by exponential enrichment (SELEX), we found a single 58-nt aptamer sequence that assumes two structu...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkp284
更新日期：2009-07-01 00:00:00
abstract：:This paper examines theoretically the effects that restraints on the tertiary structure of a superhelical DNA domain exert on the energetics of linking and the onset of conformational transitions. The most important tertiary constraint arises from the nucleosomal winding of genomic DNA in vivo. Conformational transiti...
journal_title：Nucleic acids research
pub_type： 杂志文章,评审
doi：10.1093/nar/15.23.9985
更新日期：1987-12-10 00:00:00
abstract：:By introduction of recombinant plasmids into monkey CV1 cells, we have unambiguously demonstrated that sequences entirely within the 72 bp repeat, which is located upstream of the SV40 early region, are crucial for T-antigen expression in vivo. We have also shown that a DNA fragment containing the 72 bp repeat, insert...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/9.22.6047
更新日期：1981-11-25 00:00:00
abstract：:Cell surface mannose 6-phosphate/insulin-like growth factor II receptors (M6P/IGF2R) bind and target exogenous insulin-like growth factor II (IGF2) to the prelysosomes where it is degraded. Loss of heterozygosity (LOH) for M6P/IGF2R is found in cancers, with mutational inactivation of the remaining allele. We exploite...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkj468
更新日期：2006-01-23 00:00:00
abstract：:Chromatin immunoprecipitation (ChIP) has been used to detect binding of DNA-binding proteins to sites in nuclear and mitochondrial genomes. Here, we describe a method for detecting protein-binding sites on chloroplast DNA, using modifications to the nuclear ChIP procedures. The method was developed using the lac opera...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkq413
更新日期：2010-08-01 00:00:00
abstract：:Peroxisomes are induced in Saccharomyces cerevisiae when this yeast is grown in the presence of oleate, and are repressed when glucose is supplied as the carbon source. Concomitant with this is an induction/repression of peroxisomal beta-oxidation enzymes. We are investigating the transcriptional control of acyl-CoA o...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/20.13.3495
更新日期：1992-07-11 00:00:00
abstract：:Tissue-specific gene expression is critical in understanding biological processes, physiological conditions, and disease. The identification and appropriate use of tissue-specific genes (TissGenes) will provide important insights into disease mechanisms and organ-specific therapeutic targets. To better understand the ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkx850
更新日期：2018-01-04 00:00:00
abstract：:DNA helicases are required for virtually every aspect of DNA metabolism, including replication, repair, recombination and transcription. A comprehensive description of these essential biochemical processes requires detailed understanding of helicase mechanisms. These enzymes are ubiquitous, having been identified in v...
journal_title：Nucleic acids research
pub_type： 杂志文章,评审
doi：10.1093/nar/gkl501
更新日期：2006-01-01 00:00:00
abstract：:DAP-like kinase (Dlk, also termed ZIP kinase) is a leucine zipper-containing serine/threonine-specific protein kinase with as yet unknown biological function(s). Interaction partners so far identified are either transcription factors or proteins that can support or counteract apoptosis. Thus, Dlk might be involved in ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/30.6.1408
更新日期：2002-03-15 00:00:00
abstract：:The gene for human apolipoprotein (apo) A-II has been isolated from a human genomic DNA library. The cloned fragment was approximately 14 kilobase-pair (kb) long, and extended about 9.0 kb upstream as well as 3.5 kb downstream from the apoA-II gene, which was contained within a 3.1 kb HindIII fragment of human DNA. Th...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/13.12.4597
更新日期：1985-06-25 00:00:00
abstract：:The Ensembl project (https://www.ensembl.org) annotates genomes and disseminates genomic data for vertebrate species. We create detailed and comprehensive annotation of gene structures, regulatory elements and variants, and enable comparative genomics by inferring the evolutionary history of genes and genomes. Our int...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkaa942
更新日期：2021-01-08 00:00:00
abstract：:The non-essential MGS1 gene of Saccharomyces cerevisiae is highly conserved in eukaryotes and encodes an enzyme containing both DNA-dependent ATPase and DNA annealing activities. MGS1 appears to function in post-replicational repair processes that contribute to genome stability. In this study, we identified MGS1 as a ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gki900
更新日期：2005-10-26 00:00:00
abstract：:We have previously shown that the pyrimidine oligonucleotide 5'CTTCCTCCTCT (Y11) recognizes the double-helical stem of hairpin 5'GAAGGAGGAGA-T4-TCTCCTCCTTC (h26) by triple-helix formation (1). In this paper, we report the effect on triplex formation of substituting the cytosine residues of Y11 with 5-methylcytosines (...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/19.20.5625
更新日期：1991-10-25 00:00:00
abstract：:MicroRNAs (miRNAs) are short, non-protein coding RNAs that direct the widespread phenomenon of post-transcriptional regulation of metazoan genes. The mature approximately 22-nt long RNA molecules are processed from genome-encoded stem-loop structured precursor genes. Hundreds of such genes have been experimentally val...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkn707
更新日期：2009-01-01 00:00:00
