Abstract:
:We have developed general methods for joining together, via cleavable disulfide bonds, either two unprotected polynucleotides or a polynucleotide and a peptide or protein. To join two oligonucleotides, each is first converted to an adduct in which cystamine is joined to the 5'-terminal phosphate of the oligonucleotide by a phosphoramidate bond. The adducts are mixed and reduced with dithiothreitol. The dithiothreitol is then removed by dialysis. Oxidation by atmospheric oxygen occurs to yield the required dimer. To join an oligonucleotide to a cysteine-containing peptide or protein, the 5'-cystamine oligomer is first converted to a 2'-pyridyldisulfide adduct and then reacted with an excess of the peptide or protein. If the peptide does not contain a free cysteine residue, it is first treated with iminothiolane to introduce one or more sulfhydryl groups. We have used these procedures to join a 16 mer deoxynucleotide probe and MDV-1 RNA, a substrate of Q beta RNA polymerase. This adduct hybridizes with a complementary target DNA. We have also joined a 16mer probe to peroxidase and MDV-1 RNA to human IgG. The probe-peroxidase adduct maintains enzymatic activity and the MDV-1 RNA-IgG adduct binds to a complementary anti-IgG.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Chu BC,Orgel LEdoi
10.1093/nar/16.9.3671subject
Has Abstractpub_date
1988-05-11 00:00:00pages
3671-91issue
9eissn
0305-1048issn
1362-4962journal_volume
16pub_type
杂志文章abstract::The thermal stability of the Tn10 encoded tetracycline resistance (TET) gene control region is investigated by melting studies using purified DNA restriction fragments containing various amounts of flanking sequences. In order to study the thermodynamic properties of this control region under conditions, where enough ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/10.8.2685
更新日期:1982-04-24 00:00:00
abstract::Interferons (IFN) play a pivotal role in innate immunity, orchestrating a cell-intrinsic anti-pathogenic state and stimulating adaptive immune responses. The complex interplay between the primary response to IFNs and its modulation by positive and negative feedback loops is incompletely understood. Here, we implement ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt589
更新日期:2013-09-01 00:00:00
abstract::The template requirements for efficient adenovirus DNA replication were studied in vitro in a reconstituted system with cloned DNA fragments, containing the Ad2 origin region, as templates. Replication is enhanced by nuclear factor I, a cellular protein that binds specifically to the Ad2 origin. This stimulation is sh...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/13.13.4935
更新日期:1985-07-11 00:00:00
abstract::The Trypanosoma brucei exoribonuclease, TbDSS-1, has been implicated in multiple aspects of mitochondrial RNA metabolism. Here, we investigate the role of TbDSS-1 in RNA processing and surveillance by analyzing 12S rRNA processing intermediates in TbDSS-1 RNAi cells. RNA fragments corresponding to leader sequence upst...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkm690
更新日期:2008-01-01 00:00:00
abstract::No analogous nucleoside triphosphate was found which acts as well as ATP in binding to and supporting catalysis of leucyl-tRNA synthetase from Escherichia coli MRE 600. However, there are numerous nucleotides which are able to replace ATP, but with lower efficiency. The 6-amino group of the adenine ring and the 2'-hyd...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/3.8.2067
更新日期:1976-08-01 00:00:00
abstract::A temperature-sensitive mutant of Escherichia coli affected in the alpha subunit of RNA polymerase has been investigated. Gene mapping and complementation experiments placed the mutation to temperature-sensitivity within the alpha operon at 72 min. on the bacterial chromosome. The rate of RNA synthesis in vivo and the...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.12.3445
更新日期:1990-06-25 00:00:00
abstract::MicroRNAs are small non-coding RNAs that modulate gene expression at post-transcriptional level, playing a crucial role in cell differentiation and development. Recently, some reports have shown that a limited number of mammalian microRNAs are also involved in anti-viral defense. In this study, the analysis of the hep...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr067
更新日期:2011-07-01 00:00:00
abstract::We provide a novel web service, called rQuant.web, allowing convenient access to tools for quantitative analysis of RNA sequencing data. The underlying quantitation technique rQuant is based on quadratic programming and estimates different biases induced by library preparation, sequencing and read mapping. It can tack...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq448
更新日期:2010-07-01 00:00:00
abstract::Functional genomic elements are marked by characteristic DNA and histone modification signatures. How combinatorial chromatin modification states are recognized by epigenetic reader proteins and how this is linked to their biological function is largely unknown. Here we provide a detailed molecular analysis of chromat...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw979
更新日期:2017-02-17 00:00:00
abstract::A recently reported cDNA clone coding for human promyelocytic L apoferritin, shows some differences with a liver L apoferritin cDNA. We have investigated if these differences are due to the expression of different genes or to an alternative transcription of an unique gene. In this paper we report data suggesting that ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/14.7.2863
更新日期:1986-04-11 00:00:00
abstract::We described here an approach for mapping proteins on any sequence of genomic DNA. UV-induced DNA-protein crosslinking within whole cells and the 'protein image' hybridization technique (1) were applied to test the proteins bound to different regions of the D. melanogaster hsp-70 gene. The histone H1-DNA association w...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/21.4.1031
更新日期:1993-02-25 00:00:00
abstract::Double-stranded oligonucleotides (ODNs) containing the consensus binding sequence of a transcription factor provide a rationally designed tool to manipulate gene expression at the transcriptional level by the decoy approach. However, modifications introduced into oligonucleotides to increase stability quite often do n...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh503
更新日期:2004-03-29 00:00:00
abstract::Transcript of the rbpA1 gene in Anabaena variabilis accumulates significantly at low growth temperatures below 28 degreesC. This accumulation was maximal at 16 degreesC. Accumulation of the rbpA1 transcript was completely abolished by rifampicin, but not by chloramphenicol. Photosynthesis was not required for this col...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.9.2192
更新日期:1998-05-01 00:00:00
abstract::Archival formalin-fixed, paraffin-embedded and ethanol-fixed tissues represent a potentially invaluable resource for gene expression analysis, as they are the most widely available material for studies of human disease. Little data are available evaluating whether RNA obtained from fixed (archival) tissues could produ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/30.2.e4
更新日期:2002-01-15 00:00:00
abstract::TP63, a member of the p53 gene family gene, encodes the ΔNp63 protein and is one of the most frequently amplified genes in squamous cell carcinomas (SCC) of the head and neck (HNSCC) and lungs (LUSC). Using an epiallelic series of siRNAs with intrinsically different knockdown abilities, we show that the complete loss ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw036
更新日期:2016-04-20 00:00:00
abstract::The dominant kanr marker gene plays an important role in gene disruption experiments in budding yeast, as this marker can be used in a variety of yeast strains lacking the conventional yeast markers. We have developed a loxP-kanMX-loxP gene disruption cassette, which combines the advantages of the heterologous kanr ma...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/24.13.2519
更新日期:1996-07-01 00:00:00
abstract::Sensitive detection of microsatellite instability (MSI) in tissue or liquid biopsies using next generation sequencing (NGS) has growing prognostic and predictive applications in cancer. However, the complexities of NGS make it cumbersome as compared to established multiplex-PCR detection of MSI. We present a new appro...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa1175
更新日期:2020-12-08 00:00:00
abstract::Measuring minimal residual disease in cancer has applications for prognosis, monitoring treatment and detection of recurrence. Simple sequence-based methods to detect nucleotide substitution variants have error rates (about 10-3) that limit sensitive detection. We developed and characterized the performance of MASQ (m...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa090
更新日期:2020-04-17 00:00:00
abstract::chipD is a web server that facilitates design of DNA oligonucleotide probes for high-density tiling arrays, which can be used in a number of genomic applications such as ChIP-chip or gene-expression profiling. The server implements a probe selection algorithm that takes as an input, in addition to the target sequences...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq517
更新日期:2010-07-01 00:00:00
abstract::Transfer ribonucleic acids were isolated from mitochondria of bovine heart and aminoacylated in vitro by a crude mitochondrial enzyme. Ser-tRNASerUCN and Ser-tRNASerAGY were isolated and characterized by partial sequencing. Although these tRNAs possess unique structural features not found in any bacterial tRNA, they f...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/14.18.7175
更新日期:1986-09-25 00:00:00
abstract::A challenge for large-scale siRNA loss-of-function studies is the biological pleiotropy resulting from multiple modes of action of siRNA reagents. A major confounding feature of these reagents is the microRNA-like translational quelling resulting from short regions of oligonucleotide complementarity to many different ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gku306
更新日期:2014-07-01 00:00:00
abstract::DNA polymerase beta (polbeta), a member of the X family of DNA polymerases, is the major polymerase in the base excision repair pathway. Using in vitro selection, we obtained RNA aptamers for polbeta from a variable pool of 8 x 10(12) individual RNA sequences containing 30 random nucleotides. A total of 60 individual ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkl326
更新日期:2006-05-17 00:00:00
abstract::Human ADAR3 is a catalytically inactive member of the Adenosine Deaminase Acting on RNA (ADAR) protein family, whose active members catalyze A-to-I RNA editing in metazoans. Until now, the reasons for the catalytic incapability of ADAR3 has not been defined and its biological function rarely explored. Yet, its exclusi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkz815
更新日期:2019-11-18 00:00:00
abstract::We show herein the transcription of a processed gene that originated from a spliced transcript. Recently, we isolated the human and murine homologues of the yeast chromatin protein, SPT4. The Supt4h gene is spliced normally from five exons encoded by chromosome 11. Here we show that a related sequence on chromosome 10...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.21.4960
更新日期:1998-11-01 00:00:00
abstract::The RNA-dependent RNA polymerases (RdRPs) encoded by RNA viruses represent a unique class of nucleic acid polymerases. RdRPs are essential in virus life cycle due to their central role in viral genome replication/transcription processes. However, their contribution in host adaption has not been well documented. By sol...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa1250
更新日期:2021-01-06 00:00:00
abstract::Recent evidence indicates a link between Parkinson's Disease (PD) and the expression of a-synuclein (SNCA) isoforms with different 3' untranslated regions (3'UTRs). Yet, the post-transcriptional mechanisms regulating SNCA expression are unknown. Using a large-scale in vitro /in silico screening we identified RNA-bindi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx1048
更新日期:2017-12-15 00:00:00
abstract::Nucleotide excision repair is initiated by two different damage recognition subpathways, global genome repair (GGR) and transcription-coupled repair (TCR). In GGR, XPC detects DNA lesions and recruits TFIIH via interaction with the pleckstrin homology (PH) domain of TFIIH subunit p62. In TCR, an elongating form of RNA...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx970
更新日期:2017-12-15 00:00:00
abstract::In previous studies, we reported that fractionation of HeLa cell nuclear extracts on glycerol gradients revealed an endogenous ∼10S particle that contained galectin-3 and U1 snRNP and this particle was sufficient to load the galectin polypeptide onto a pre-mRNA substrate. We now document that this interaction between ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw303
更新日期:2016-07-27 00:00:00
abstract::Mechanisms that preserve genome integrity are highly important during the normal life cycle of human cells. Loss of genome protective mechanisms can lead to the development of diseases such as cancer. Checkpoint kinases function in the cellular surveillance pathways that help cells to cope with DNA damage. Importantly...
journal_title:Nucleic acids research
pub_type: 杂志文章,评审
doi:10.1093/nar/gkr697
更新日期:2012-01-01 00:00:00
abstract::We have synthesized the tetradecamer GAGCXGATAACAAG containing a part of the sequence of the lactose operator. A guanine base in the sequence is replaced by the adduct of the carcinogen 2-acetylaminofluorene with guanine. Under the standard conditions of de-protection, the fluorene moiety is lost, leaving behind a gua...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/11.15.5093
更新日期:1983-08-11 00:00:00