Abstract:
:Kinetically monitored, reverse transcriptase-initiated PCR (kinetic RT-PCR, kRT-PCR) is a novel application of kinetic PCR for high throughput transcript quantitation in total cellular RNA. The assay offers the simplicity and flexibility of an enzyme assay with distinct advantages over DNA microarray hybridization and SAGE technologies for certain applications. The reproducibility, sensitivity and accuracy of the kRT-PCR were assessed for yeast transcripts previously quantitated by a variety of methods including SAGE analysis. Changes in transcript levels between different genetic or physiological cell states were reproducibly quantitated with an accuracy of +/-20%. The assay was sufficiently sensitive to quantitate yeast transcripts over a range of more than five orders of magnitude, including low abundance transcripts encoding cell cycle and transcriptional regulators.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Kang JJ,Watson RM,Fisher ME,Higuchi R,Gelfand DH,Holland MJdoi
10.1093/nar/28.2.e2keywords:
subject
Has Abstractpub_date
2000-01-15 00:00:00pages
e2issue
2eissn
0305-1048issn
1362-4962pii
gnd002journal_volume
28pub_type
杂志文章abstract::The unwinding of supercoiled phi X174 RFI DNA induced by the tumorigenic (+) and non-tumorigenic (-) enantiomers of trans-7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BPDE) has been investigated by agarose slab-gel and ethidium titration tube gel electrophoresis. The differences in adduct conformat...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/20.23.6167
更新日期:1992-12-11 00:00:00
abstract::We describe an interactive computer program (ANALYSEQ) that is used from a simple graphics terminal. The main purpose of the program is to determine the function of nucleic acid sequences but it also offers the simpler listing, searching and counting options. It contains methods to locate genes by looking for the effe...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/12.1part2.521
更新日期:1984-01-11 00:00:00
abstract::Despite the large number of noncoding RNAs in human genome and their roles in many diseases include cancer, we know very little about them due to lack of structural clues. The centerpiece of the structural clues is the full RNA base-pairing structure of secondary and tertiary contacts that can be precisely obtained on...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkz1192
更新日期:2020-02-20 00:00:00
abstract::Computational modelling of mammalian cell cycle regulation is a challenging task, which requires comprehensive knowledge on many interrelated processes in the cell. We have developed a web-based integrated database on cell cycle regulation in mammals in normal and pathological states (Cyclonet database). It integrates...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkl912
更新日期:2007-01-01 00:00:00
abstract::Aspartic acid residues comprising the -D-(aa) n -Y-L-D-D- DNA polymerase active site triad of reverse transcriptase from the Saccharomyces cerevisiae long terminal repeat-retrotransposon Ty3 (Asp151, Asp213 and Asp214) were evaluated via site-directed mutagenesis. An Asp151-->Glu substitution showed a dramatic decreas...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gki150
更新日期:2005-01-12 00:00:00
abstract::We determined the structure around the gap region between 28S alpha and 28S beta rRNA of Bombyx mori, a lepidopteran insect, to know the introduction mechanism of the hidden break, an interruption of the phosphodiester bond specific to the 28S rRNA of protostomes. Sequence analysis and S1 nuclease mapping suggested th...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/14.16.6393
更新日期:1986-08-26 00:00:00
abstract::Transcriptomics (at the level of single cells, tissues and/or whole organisms) underpins many fields of biomedical science, from understanding the basic cellular function in model organisms, to the elucidation of the biological events that govern the development and progression of human diseases, and the exploration o...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq667
更新日期:2010-09-01 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::Gene duplication is an important process in evolution. The availability of genome sequences of a number of organisms has made it possible to conduct comprehensive searches for duplicated genes enabling informative studies of their evolution. We have established the FGF (Fishing Gene Family) program to efficiently sear...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkm426
更新日期:2007-07-01 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::Conventional approaches to predict transcriptional regulatory interactions usually rely on the definition of a shared motif sequence on the target genes of a transcription factor (TF). These efforts have been frustrated by the limited availability and accuracy of TF binding site motifs, usually represented as position...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gks1286
更新日期:2013-02-01 00:00:00
abstract::MicroRNAs (miRNA) are short single-stranded RNA molecules derived from hairpin-forming precursors that play a crucial role as post-transcriptional regulators in eukaryotes and viruses. In the past years, many microRNA target genes (MTGs) have been identified experimentally. However, because of the high costs of experi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw1085
更新日期:2017-01-25 00:00:00
abstract::A segment of 1160 nucleotides of the FMDV genome has been sequenced using three overlapping fragments of cloned cDNA from FMDV strain O1K. This sequence contains the coding sequence for the viral capsid protein VP1 as shown by its homology to known and newly determined amino acid sequences from this man antigenic poly...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/9.8.1919
更新日期:1981-04-24 00:00:00
abstract::The high-throughput sequencing of nuclease-protected mRNA fragments bound to ribosomes, a technique known as ribosome profiling, quantifies the relative frequencies with which different regions of transcripts are translated. This technique has revealed novel translation initiation sites with unprecedented scope and ha...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv200
更新日期:2015-04-20 00:00:00
abstract::The Escherichia coli AlkB protein catalyzes the direct reversal of alkylation damage to DNA; primarily 1-methyladenine (1mA) and 3-methylcytosine (3mC) lesions created by endogenous or environmental alkylating agents. AlkB is a member of the non-heme iron (II) alpha-ketoglutarate-dependent dioxygenase superfamily, whi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkm1031
更新日期:2007-01-01 00:00:00
abstract::The HIV RT and Protease Sequence Database is an online relational database that catalogs evolutionary and drug-related human immunodeficiency virus (HIV) reverse transcriptase (RT) and protease sequence variation (http://hivdb.stanford.edu). The database contains a compilation of nearly all published HIV RT and protea...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/28.1.346
更新日期:2000-01-01 00:00:00
abstract::The wobble uridine of certain bacterial and mitochondrial tRNAs is modified, at position 5, through an unknown reaction pathway that utilizes the evolutionarily conserved MnmE and GidA proteins. The resulting modification (a methyluridine derivative) plays a critical role in decoding NNG/A codons and reading frame mai...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkp762
更新日期:2009-11-01 00:00:00
abstract::Proline-rich protein (PRP) gene MP6 was isolated from a mouse BALB/c genomic DNA library in lambda EMBL3, characterised by hybridisation and restriction mapping and the promoter region, from -162 to +72 around the PRP consensus cap-site, was sequenced. In gel shift assays this region formed complexes C1 and C2 with pa...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/19.19.5205
更新日期:1991-10-11 00:00:00
abstract::The anaerobically-regulated nirB promoter was used to express heterologous genes in Escherichia coli. Under anaerobic conditions the promoter was able to express tetanus toxin fragment C at approximately 20% total cell protein (tcp) and the Bordetella pertussis antigen pertactin at greater than 30% tcp. These levels a...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/19.11.2889
更新日期:1991-06-11 00:00:00
abstract::The sequence requirements for transcriptional stimulation of the adenovirus major late promoter (MLP) by the products of the early transcription unit Ela and by the replication of viral DNA were analyzed by in vitro transcription. Sequences upstream of +33 are involved in the moderate Ela-responsiveness of the MLP, wh...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/16.9.3771
更新日期:1988-05-11 00:00:00
abstract::The recent outbreak of COVID-19 has generated an enormous amount of Big Data. To date, the COVID-19 Open Research Dataset (CORD-19), lists ∼130,000 articles from the WHO COVID-19 database, PubMed Central, medRxiv, and bioRxiv, as collected by Semantic Scholar. According to LitCovid (11 August 2020), ∼40,300 COVID19-re...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa969
更新日期:2021-01-08 00:00:00
abstract::Short interfering RNAs (siRNAs) are valuable reagents for sequence-specific inhibition of gene expression via the RNA interference (RNAi) pathway. Although it has been proposed that the relative thermodynamic stability at the 5'-ends of siRNAs plays a crucial role in siRNA strand selection, we demonstrate here that a ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkn584
更新日期:2008-10-01 00:00:00
abstract::E. coli genes for tRNAasp and tRNAtrp are transcribed by eukaryotic RNA polymerase III upon injection into nuclei of Xenopus oocytes. This indicates that some bacterial tRNA genes contain RNA polymerase III promoter sequences. We discuss evolutionary aspects of this finding. ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/10.22.7153
更新日期:1982-11-25 00:00:00
abstract::Interactions between subunits in the Saccharomyces cerevisiae ribosome are mediated by universal and eukaryote-specific intersubunit bridges. Universal bridges are positioned close to the ribosomal functional centers, while eukaryote-specific bridges are mainly located on the periphery of the ribosome. Two bridges, eB...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky1083
更新日期:2019-01-10 00:00:00
abstract::Multi-modal interactions are frequently observed in intrinsically disordered regions (IDRs) of proteins upon binding to their partners. In many cases, post-translational modifications in IDRs are accompanied by coupled folding and binding. From both molecular simulations and biochemical experiments with mutational stu...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx1297
更新日期:2018-03-16 00:00:00
abstract::This review presents detailed information about the structure of triplet repeat RNA and addresses the simple sequence repeats of normal and expanded lengths in the context of the physiological and pathogenic roles played in human cells. First, we discuss the occurrence and frequency of various trinucleotide repeats in...
journal_title:Nucleic acids research
pub_type: 杂志文章,评审
doi:10.1093/nar/gkr729
更新日期:2012-01-01 00:00:00
abstract::Eukaryotic cells possess several distinct mismatch repair pathways. A mismatch can be introduced in retroviral double-stranded DNA by a pre-existing mutation within the primer binding site (PBS) of the viral RNA genome. In order to evaluate mismatch repair of retroviral double-stranded DNA, Moloney leukemia virus (MLV...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/28.12.2302
更新日期:2000-06-15 00:00:00
abstract::The adenovirus late region 1 (L1) represents an example of an alternatively spliced gene where one 5' splice site is spliced to two alternative 3' splice sites, to produce two mRNAs; the 52,55K and IIIa mRNAs, respectively. Accumulation of the L1 mRNAs is temporally regulated during the infectious cycle. Thus, the pro...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/22.3.332
更新日期:1994-02-11 00:00:00
abstract::The Est3 subunit of yeast telomerase, which adopts a predicted OB-fold, is essential for telomere replication. To assess the possible contributions that Est3 might make to enzyme catalysis, we compared telomerase activity from wild type and est3-Delta strains of Saccharomyces castellii, which revealed that loss of the...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkp1173
更新日期:2010-04-01 00:00:00
abstract::The stable intermediates formed in the reaction of cytosine, cytidine and adenosine with chloracetaldehyde were isolated. The -CH2CH/OH/- bridge between the exo and endo nitrogen atoms of the parent base was found in these compounds by means of PMR spectroscopy. Their acid-induced dehydration resulted in formation of ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/5.3.789
更新日期:1978-03-01 00:00:00