Abstract：
:In vitro enzymatic amplification of nucleic acids by PCR or other techniques is a very sensitive method to detect rare DNA segments. We present here a protocol that allows the rapid, sensitive and precise quantification of DNA molecules using PCR amplification run to saturation. The DNA (or cDNA) to be assayed is co-amplified with known amounts of an internal standard DNA. We show that the latter must be almost identical to the assayed DNA, otherwise quantification at the plateau is unreliable. The read-out of the amplification involves one or two additional oligonucleotides. Using fluorescent oligonucleotides as primers in run-off reactions together with an automated DNA sequencer, we could measure the level of expression of several genes, like the murine MHC class I H-2Kd or a specific T cell receptor beta chain transcript in the course of an immunization. mRNA levels were normalized by measuring in a similar manner the number of transcripts encoding the housekeeping gene HPRT. Finally, our procedure might allow the rapid analysis of a large number of samples at the same time, as illustrated by the simultaneous analysis of the mRNAs encoding the CD4 and CD8 murine T cell markers.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Pannetier C,Delassus S,Darche S,Saucier C,Kourilsky Pdoi
10.1093/nar/21.3.577subject
Has Abstractpub_date
1993-02-11 00:00:00pages
577-83issue
3eissn
0305-1048issn
1362-4962journal_volume
21pub_type
杂志文章abstract：:Transcription initiation complexes formed by bacterial RNA polymerases (RNAPs) exhibit dramatic species-specific differences in stability, leading to different strategies of transcription regulation. The molecular basis for this diversity is unclear. Promoter complexes formed by RNAP from Thermus aquaticus (Taq) are c...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gks973
更新日期：2012-12-01 00:00:00
abstract：:Bulk replicative DNA synthesis in eukaryotes is highly accurate and efficient, primarily because of two DNA polymerases (Pols): Pols δ and ε. The high fidelity of these enzymes is due to their intrinsic base selectivity and proofreading exonuclease activity which, when coupled with post-replication mismatch repair, he...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkq1034
更新日期：2011-03-01 00:00:00
abstract：:Lambda exonuclease processively degrades one strand of double-stranded DNA (dsDNA) in the 5"-3" direction. To understand the mechanism through which this enzyme generates high processivity we are analyzing the first step in the reaction, namely the interaction of lambda exonuclease with the ends of substrate DNA. Endo...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/27.15.3057
更新日期：1999-08-01 00:00:00
abstract：:We created a single cell sorting system to screen for enzyme activity in Escherichia coli producing 3,4 dihydroxy benzoate (34DHB). To do so, we engineered a transcription factor regulon controlling the expression of green fluorescent protein (GFP) for induction by 34DHB. An autoregulated transcription factor, pcaU, w...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gku444
更新日期：2014-07-01 00:00:00
abstract：:Toxicogenomics focuses on assessing the safety of compounds using gene expression profiles. Gene expression signatures from large toxicogenomics databases are expected to perform better than small databases in identifying biomarkers for the prediction and evaluation of drug safety based on a compound's toxicological m...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gku955
更新日期：2015-01-01 00:00:00
abstract：:We have used paired-end sequencing of yeast nucleosomal DNA to obtain accurate genomic maps of nucleosome positions and occupancies in control cells and cells treated with 3-aminotriazole (3AT), an inducer of the transcriptional activator Gcn4. In control cells, 3AT-inducible genes exhibit a series of distinct nucleos...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkr643
更新日期：2011-12-01 00:00:00
abstract：:Structural information on interacting proteins is important for understanding life processes at the molecular level. Genome-wide docking database is an integrated resource for structural studies of protein-protein interactions on the genome scale, which combines the available experimental data with models obtained by ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkp944
更新日期：2010-01-01 00:00:00
abstract：:Nucleotide sequences of three cloned restriction fragments of Tetrahymena mtDNA which showed hybridization with mitochondrial tRNA have been determined. EcoRI fragment 5 (4.1 kbp) contains the tRNAphe gene sequence with anticodon GAA; Hind III fragment 6 (2.0 kbp) the tRNAhis with anticodon GTG; and EcoRI fragment 7 (...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/13.9.3273
更新日期：1985-05-10 00:00:00
abstract：:Fifteen years elapsed between the discovery of the double helix (1953) and the first DNA sequencing (1968). Modern DNA sequencing began in 1977, with development of the chemical method of Maxam and Gilbert and the dideoxy method of Sanger, Nicklen and Coulson, and with the first complete DNA sequence (phage X174), whi...
journal_title：Nucleic acids research
pub_type： 历史文章,杂志文章
doi：10.1093/nar/gkm688
更新日期：2007-01-01 00:00:00
abstract：:Insertion of microRNA target sequences into the flavivirus genome results in selective tissue-specific attenuation and host-range restriction of live attenuated vaccine viruses. However, previous strategies for miRNA-targeting did not incorporate a mechanism to prevent target elimination under miRNA-mediated selective...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkw061
更新日期：2016-04-20 00:00:00
abstract：:In plants, small interfering RNAs (siRNAs) can trigger a silencing signal that may spread within a tissue to adjacent cells or even systemically to other organs. Movement of the signal is initially limited to a few cells, but in some cases the signal can be amplified and travel over larger distances. How far silencing...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkq1240
更新日期：2011-04-01 00:00:00
abstract：:Eight base analogs were tested as third strand residues in otherwise homopyrimidine strands opposite each of the 'direct' (A.T and G.C) and 'inverted' (T.A and C.G) Watson-Crick base pairs, using UV melting profiles to assess triplex stability. The target duplexes contained 20 A.T base pairs and a central test base pa...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/27.23.4632
更新日期：1999-12-01 00:00:00
abstract：:Endonuclease III from Escherichia coli, yeast (yNtg1p and yNtg2p) and human and E.coli endonuclease VIII have a wide substrate specificity, and recognize oxidation products of both thymine and cytosine. DNA containing single dihydrouracil (DHU) and tandem DHU lesions were used as substrates for these repair enzymes. I...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/29.2.407
更新日期：2001-01-15 00:00:00
abstract：:We report thermodynamic values for binding of the guanosine nucleophile to the ribozyme derived from the Anabaena group I intron, and find that they are similar to those measured previously for the structurally distinct Tetrahymena ribozyme. The free energy of binding guanosine 5'-monophosphate (pG) at 30 degrees C is...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/24.19.3722
更新日期：1996-10-01 00:00:00
abstract：:We attempted to use the polymerase chain reaction (PCR) to monitor in vitro recombination in a plasmid containing directly repeated sequences. Some of the plasmid preparations which had not been exposed to recombination conditions were however found to behave in the PCR test as if they had undergone homologous recombi...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/19.9.2423
更新日期：1991-05-11 00:00:00
abstract：:We describe a new exonuclease-based method for joining and/or constructing two or more DNA molecules. DNA fragments containing ends complementary to those of a vector or another independent molecules were generated by the polymerase chain reaction. The 3' ends of these molecules as well as the vector DNA were then rec...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/21.8.1889
更新日期：1993-04-25 00:00:00
abstract：:The discontinuous synthesis of the complementary strands of polyoma DNA in isolated nuclei has been studied by hybridization techniques. The relative amounts of the newly synthesized complementary strands were compared by separately annealing them to denatured HpaII restriction fragments. In every case an excess (1.4-...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/4.5.1449
更新日期：1977-01-01 00:00:00
abstract：:We have isolated an eight kilobase fragment of Bacillus subtilis DNA by specific integration and excision of a plasmid containing a sequence adjacent to ribosomal operon rrn O. The genetic locus of the cloned fragment was verified by linkage of the integrated vector to nearby genetic markers using both transduction an...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/12.15.6307
更新日期：1984-08-10 00:00:00
abstract：:In many organisms (e.g., gram-positive eubacteria) Gin-tRNA is not formed by direct glutaminylation of tRNAGln but by a specific transamidation of Glu-tRNAGln. We wondered whether a similar transamidation pathway also operates in the formation of Asn-tRNA in these organisms. Therefore we tested in S-100 preparations o...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/24.14.2648
更新日期：1996-07-15 00:00:00
abstract：:Polyaromatic molecules with amino chain substituents, upon binding with DNA, selectively catalyze exchange of the A.T base pair protons with bulk water protons. The amine-catalyzed exchange is mediated by compounds which are A.T and G.C base sequence specific, intercalators, and outside binders. A mechanism for the se...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/15.20.8511
更新日期：1987-10-26 00:00:00
abstract：:Signal transducer and activator of transcription 6 (STAT6) is a regulator of transcription for interleukin-4 (IL-4)-induced genes. The ability of STAT6 to activate transcription depends on functional interaction with other transcription factors and coactivators. We have characterized the mechanism of STAT6-mediated tr...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkl539
更新日期：2006-01-01 00:00:00
abstract：:We have previously shown that hedamycin, a GC-rich DNA-binding antitumor agent, downregulates survivin transcription (Wu et al. (2005) Molecular mechanism of inhibition of survivin transcription by the GC-rich sequence selective DNA-binding antitumor agent, hedamycin: evidence of survivin downregulation associated wit...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkm149
更新日期：2007-01-01 00:00:00
abstract：:The distribution of 7-methylguanine in the families of repetitive and unique sequences of rat liver chromatin DNA has been studied using the technique of DNA-DNA reassociation. Rats were injected with di[14C]methylnitrosamine and chromatin DNA was prepared 3 h later. The distribution of 7-methylguanine was found to be...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/5.9.3357
更新日期：1978-09-01 00:00:00
abstract：:Cellular RNAs that do not function as messenger RNAs (mRNAs), transfer RNAs (tRNAs) or ribosomal RNAs (rRNAs) comprise a diverse class of molecules that are commonly referred to as non-protein-coding RNAs (ncRNAs). These molecules have been known for quite a while, but their importance was not fully appreciated until ...
journal_title：Nucleic acids research
pub_type： 杂志文章,评审
doi：10.1093/nar/gkj469
更新日期：2006-01-25 00:00:00
abstract：:Enhancer-dependent transcription involving the promoter specificity factor σ(54) is widely distributed amongst bacteria and commonly associated with cell envelope function. For transcription initiation, σ(54)-RNA polymerase yields open promoter complexes through its remodelling by cognate AAA+ ATPase activators. Since...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkv597
更新日期：2015-09-03 00:00:00
abstract：:Endonucleases that generate double-strand breaks in DNA often possess two identical subunits related by rotational symmetry, arranged so that the active sites from each subunit act on opposite DNA strands. In contrast to many endonucleases, Type IIP restriction enzyme BcnI, which recognizes the pseudopalindromic seque...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkr588
更新日期：2011-11-01 00:00:00
abstract：:Acinetobacter sp. strain ADP1 is a nutritionally versatile soil bacterium closely related to representatives of the well-characterized Pseudomonas aeruginosa and Pseudomonas putida. Unlike these bacteria, the Acinetobacter ADP1 is highly competent for natural transformation which affords extraordinary convenience for ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkh910
更新日期：2004-10-28 00:00:00
abstract：:The availability of protein fluorophores with appropriate spectral properties has made it possible to employ fluorescence resonance energy transfer (FRET) to assess interactions between three proteins microscopically. Flow cytometry offers excellent sensitivity, effective signal separation and the capacity to assess a...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gni057
更新日期：2005-04-01 00:00:00
abstract：:We have cloned and sequenced the lasR gene, which is involved in the transcriptional activation of several pathogenic factors, from Pseudomonas aeruginosa IFO3455 and PA103. These clones were predicted to be an open reading frame of 239 amino acids as reported for the PAO1 strain. There is only a single base change re...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/22.18.3706
更新日期：1994-09-11 00:00:00
abstract：:Dinucleoside diphosphates of the general type pGpN have been prepared enzymatically using ribonuclease N1. Alkylated uridines or cytidines, which are products of carcinogens acting on nucleic acids, were tested in dinucleoside diphosphates for their ability to stimulate the binding of Ala- or Val-tRNA to ribosomes. O2...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/6.4.1709
更新日期：1979-04-01 00:00:00
