Abstract：
:Inability of T7 RNA polymerase to processively transcribe higher eukaryotic chromatin is interpreted as a correlate of its reported inhibition by nucleosomes on reconstituted templates in vitro . We used chromosomally integrated reporter cassettes to examine features of T7 transcription in a lower eukaryotic system. Luciferase reporters were targeted to rDNA in transgenic Trypanosoma brucei stably expressing the phage polymerase. Because trypanosome mRNAs are capped by RNA splicing in trans , T7 transcription could be gauged by luciferase activity. In contrast to findings from higher eukaryotes, T7 transcription is vigorous and processive on chromatin templates in T.brucei , surpassing levels achieved with endogenous promoters, including those recruiting RNA polymerase I. This may be a reflection of intrinsic differences in chromatin structure between differently evolved eukaryotes or of an integration site that is exceptionally permissive for T7 transcription due to a local accessible chromatin conformation. T7 transcription could be manipulated to achieve different levels of constitutive expression, through the use of promoter mutations. Moreover, T7 initiation could be regulated by the prokaryotic Tet repressor and elongation halted by T7 terminator sequences. We have exploited these features to construct a robust inducible expression system, whose utility potentially extends to other trans -splicing organisms.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Wirtz E,Hoek M,Cross GAdoi
10.1093/nar/26.20.4626subject
Has Abstractpub_date
1998-10-15 00:00:00pages
4626-34issue
20eissn
0305-1048issn
1362-4962pii
gkb742journal_volume
26pub_type
杂志文章abstract：:The functionalisation of oligodeoxynucleotides and oligoribonucleotides by incorporation of 2'-amido-2'-deoxyribonucleosides, possibly containing a reporter group via the 2'-amido bond, was examined. Therefore 2'-acetamido-ribonucleosides containing a small methyl group at the 2'-amido bond were synthesized as model c...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/23.1.51
更新日期：1995-01-11 00:00:00
abstract：:The polymerase encoded by human hepatitis B virus, which has reverse transcriptase and RNase H activity, binds to its pregenomic RNA template in a two-step process involving a terminal redundancy. Both first strand and second strand DNA synthesis involve primer translocation and second strand synthesis involves a temp...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/24.17.3295
更新日期：1996-09-01 00:00:00
abstract：:An intronic hexanucleotide UGCAUG has been shown to play a critical role in the regulation of tissue-specific alternative splicing of pre-mRNAs in a wide range of tissues. Vertebrate Fox-1 has been shown to bind to this element, in a highly sequence-specific manner, through its RNA recognition motif (RRM). In mammals,...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gki338
更新日期：2005-04-11 00:00:00
abstract：:An improved contact hybridization technique for the analysis of labelled RNA species is presented. The method combines high sensitivity of detection with the high resolution of polyacrylamide gel electrophoresis and should be especially useful for the characterization of transient RNA precursor molecules. Its applicat...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/6.9.2963
更新日期：1979-07-11 00:00:00
abstract：:In vivo and in vitro RNA splicing experiments have demonstrated that the intron splicing machineries are not interchangeable in all organisms. These differences have prevented the efficient in vivo expression of monocot genes containing introns in dicot plants and the in vitro excision of some plant introns in HeLa ce...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/16.14.7159
更新日期：1988-07-25 00:00:00
abstract：:PipeAlign is a protein family analysis tool integrating a five step process ranging from the search for sequence homologues in protein and 3D structure databases to the definition of the hierarchical relationships within and between subfamilies. The complete, automatic pipeline takes a single sequence or a set of sequ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkg518
更新日期：2003-07-01 00:00:00
abstract：:We have analysed DNA modification in a HapII site (CCGG) present in the major intron of the discontinuous rabbit beta-globin gene. In most somatic tissues, including erythroid and non-erythroid tissues, about 50% of the DNA is resistant to cleavage at this site by HapII, though 100% cleavage is found with the isoschiz...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/5.12.4631
更新日期：1978-12-01 00:00:00
abstract：:Nucleotide excision repair (NER) mechanism is the major pathway responsible for the removal of a large variety of bulky lesions from the genome. Two different NER subpathways have been identified, i.e. the transcription-coupled and the global genome repair pathways. For DNA-damage induced by ultraviolet light both tra...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/24.23.4653
更新日期：1996-12-01 00:00:00
abstract：:We have identified and sequenced two members of a chicken middle repetitive DNA sequence family. By reassociation kinetics, members of this family (termed CRl) are estimated to be present in 1500-7000 copies per chicken haploid genome. The first family member sequenced (CRlUla) is located approximately 2 kb upstream f...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/9.20.5383
更新日期：1981-10-24 00:00:00
abstract：:Post-translational modifications contribute to the spliceosome dynamics by facilitating the physical rearrangements of the spliceosome. Here, we report USP15, a deubiquitinating enzyme, as a regulator of protein-protein interactions for the spliceosome dynamics. We show that PRP31, a component of U4 snRNP, is modified...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkw1365
更新日期：2017-05-05 00:00:00
abstract：:The paper describes an improved method for the synthesis of oligodeoxyribonucleotides using phosphoramidite chemistry. Our procedure relies on novel phosphoramidite intermediates, the deoxyribonucleoside-3'-morpholine-methoxyphosphins. These compounds are extremely stable and can be purified readily. Condensation reac...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/11.9.2575
更新日期：1983-05-11 00:00:00
abstract：:We have constructed simple test-plasmids to study transcriptional enhancers in yeast. In this paper the reporter-gene is a plasmid borne deletion-substitution derivative (his-del4) of the Saccharomyces cerevisiae HIS3 gene in which the native promoter has been replaced by a dormant, susceptible pseudopromoter. We inve...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/16.18.8847
更新日期：1988-09-26 00:00:00
abstract：:A simple, reproducible affinity chromatography method has been adapted for separation of high molecular weight supercoiled circular molecules from mammalian cells. Electron microscopic analysis of EB viral DNA obtained by this method, from the non-producer Burkitt's lymphoma line Raji, revealed monomer-sized viral mol...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/15.24.10345
更新日期：1987-12-23 00:00:00
abstract：:RNA editing by adensosine deaminases is a widespread mechanism to alter genetic information in metazoa. In addition to modifications in non-coding regions, editing contributes to diversification of protein function, in analogy to alternative splicing. However, although splicing programs respond to external signals, fa...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gks1045
更新日期：2013-01-01 00:00:00
abstract：:Hidden Markov models (HMMs) and their variants are widely used in Bioinformatics applications that analyze and compare biological sequences. Designing a novel application requires the insight of a human expert to define the model's architecture. The implementation of prediction algorithms and algorithms to train the m...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkp662
更新日期：2009-11-01 00:00:00
abstract：:The cancer drug daunomycin is an effective inhibitor of the B leads to Z transition in poly d(G-C) in 4 M NaCl. Both the rate and extent of the B leads to Z transition are decreased by the drug, as judged by equilibrium and kinetic studies. Daunomycin can, under some conditions, convert Z form DNA back to B form. Drug...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/11.23.8485
更新日期：1983-12-10 00:00:00
abstract：:The web software CHOP was developed to visualize the 'wobbling' in the third codon position of aligned DNA sequences. The simple features of this tool allow users to easily find regions suspected of containing coding sequences (CDSs). The program also allows visualization of the nucleotide diversity between two genomi...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkh448
更新日期：2004-07-01 00:00:00
abstract：:DNA, isolated from age-synchronous senescent populations of Caenorhabditis elegans has been quantitatively and qualitatively analyzed for the presence of 5-methylcytosine. High performance liquid chromatography on two wild-type and several mutant strains of C. elegans failed to detect any 5-methylcytosine. The restric...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/14.16.6711
更新日期：1986-08-26 00:00:00
abstract：:We found previously that the Chlamydomonas HSP70A promoter counteracts transcriptional silencing of downstream promoters in a transgene setting. To elucidate the underlying mechanisms, we analyzed chromatin state and transgene expression in transformants containing HSP70A-RBCS2-ble (AR-ble) constructs harboring deleti...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkt224
更新日期：2013-05-01 00:00:00
abstract：:The endonuclease activity within the influenza virus cap-snatching process is a proven therapeutic target. The anti-influenza drug baloxavir is highly effective, but is associated with resistance mutations that threaten its clinical efficacy. The endonuclease resides within the N-terminal domain of the PA subunit (PAN...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkaa1294
更新日期：2021-01-19 00:00:00
abstract：:In order to characterize structure-function relationships, the kinetic behavior of chimeric RNA/DNA ribozyme was compared with that of all RNA ribozyme. Determined kcat values were proven to represent the chemical-cleavage step and not the product-dissociation step. In agreement with the finding by Dahm and Uhlenbeck ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/21.24.5656
更新日期：1993-12-11 00:00:00
abstract：:Pluripotency, the ability of embryonic stem cells to differentiate into specialized cell types, is determined by ESC-specific gene regulators such as transcription factors and chromatin modification factors. It is not well understood how ESCs are poised for differentiation, however, and methods are needed for prognosi...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkx752
更新日期：2017-10-13 00:00:00
abstract：:RNase III enzymes cleave double stranded (ds)RNA. This is an essential step for regulating the processing of mRNA, rRNA, snoRNA and other small RNAs, including siRNA and miRNA. Arabidopsis thaliana encodes nine RNase III: four DICER-LIKE (DCL) and five RNASE THREE LIKE (RTL). To better understand the molecular functio...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkx820
更新日期：2017-11-16 00:00:00
abstract：:Transcription initiation is a major step in gene regulation for all organisms. In bacteria, the promoter DNA is first recognized by RNA polymerase (RNAP) to yield an initial closed complex. This complex subsequently undergoes conformational changes resulting in DNA strand separation to form a transcription bubble and ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkx1146
更新日期：2018-01-25 00:00:00
abstract：:PrimPol is a primase-polymerase involved in nuclear and mitochondrial DNA replication in eukaryotic cells. Although PrimPol is predicted to possess an archaeo-eukaryotic primase and a UL52-like zinc finger domain, the role of these domains has not been established. Here, we report that the proposed zinc finger domain ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gku214
更新日期：2014-05-01 00:00:00
abstract：:New linker phosphoramidite reagents containing a cleavable 3'-ester linkage are used for attaching the first nucleoside to the surface of a solid- phase support. Inexpensive, underivatized amino supports, such as long chain alkylamine controlled-pore glass, can serve as universal supports. No modifications to phosphor...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkh222
更新日期：2004-01-29 00:00:00
abstract：:The gene co-expression analysis toolbox (GeneCAT) introduces several novel microarray data analyzing tools. First, the multigene co-expression analysis, combined with co-expressed gene networks, provides a more powerful data mining technique than standard, single-gene co-expression analysis. Second, the high-throughpu...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkn292
更新日期：2008-07-01 00:00:00
abstract：:Nuclear receptors for the thyroid hormone and vitamin A and D cooperate with the retinoid X receptor (RXR) in activating the transcription. Although the hormone response elements for these receptors have been proposed in which spacing of the direct repeated motifs determine the specificity (so called 3-4-5 rule), vita...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/22.15.2902
更新日期：1994-08-11 00:00:00
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：:LRPPRC is a protein that has attracted interest both for its role in post-transcriptional regulation of mitochondrial gene expression and more recently because numerous mutated variants have been characterized as causing severe infantile mitochondrial neurodegeneration. LRPPRC belongs to the pentatricopeptide repeat (...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkw575
更新日期：2016-08-19 00:00:00
