Abstract:
:Strand Displacement Amplification (SDA) is an isothermal, in vitro method of amplifying a DNA target sequence prior to detection [Walker et al (1992) Nucleic Acids Res., 20, 1691-1693]. Here we describe a multiplex form of SDA that allows two target sequences and an internal amplification control to be co-amplified by a single pair of primers after common priming sequences are spontaneously appended to the ends of target fragments. Multiplex SDA operates at a single temperature, under the same simple protocol previously developed for single-target SDA. We applied multiplex SDA to co-amplification of a target sequence (IS6110) that is specific to members of the Mycobacterium tuberculosis-complex and a target (16S ribosomal gene) that is common to most clinically relevant species of mycobacteria. Both targets are amplified 10(8)-fold during a 2 hour, single temperature incubation. The relative sensitivity of the system was evaluated across a number of clinically relevant mycobacteria and checked for crossreactivity against organisms that are closely related to mycobacteria.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Walker GT,Nadeau JG,Spears PA,Schram JL,Nycz CM,Shank DDdoi
10.1093/nar/22.13.2670subject
Has Abstractpub_date
1994-07-11 00:00:00pages
2670-7issue
13eissn
0305-1048issn
1362-4962journal_volume
22pub_type
杂志文章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::The STE2 gene of the yeast Saccharomyces cerevisiae encodes a component of the receptor for the oligopeptide pheromone alpha-factor. We have cloned and determined the nucleotide sequence of the STE2 gene. A sequence involved in the control of cell-type expression of the STE2 gene was found 5' of an open reading frame ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/13.23.8463
更新日期:1985-12-09 00:00:00
abstract::Alternative splicing and polyadenylation were observed pervasively in eukaryotic messenger RNAs. These alternative isoforms could either be consequences of physiological regulation or stochastic noise of RNA processing. To quantify the extent of stochastic noise in splicing and polyadenylation, we analyzed the alterna...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gks1271
更新日期:2013-02-01 00:00:00
abstract::Bacillus subtilis RecN appears to be an early detector of breaks in double-stranded DNA. In vivo, RecN forms discrete nucleoid-associated structures and in vitro exhibits Mg2+-dependent single-stranded (ss) DNA binding and ssDNA-dependent ATPase activities. In the presence of ATP or ADP, RecN assembles to form large n...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gki533
更新日期:2005-04-22 00:00:00
abstract::A new series of DNA bis-intercalators is reported in which acridine moieties are connected by rigid and extended pyridine-based linkers of varied length. Cross-linking of DNA by bis-intercalation is inferred from the unwinding and folding of linear DNA induced by the compounds; after ligation and removal of the bis-in...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/20.5.983
更新日期:1992-03-11 00:00:00
abstract::The E. coli protein StpA has RNA annealing and strand displacement activities and it promotes folding of RNAs by loosening their structures. To understand the mode of action of StpA, we analysed the relationship of its RNA chaperone activity to its RNA-binding properties. For acceleration of annealing of two short RNA...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkl1143
更新日期:2007-01-01 00:00:00
abstract::The control of messenger RNA (mRNA) function by micro RNAs (miRNAs) in animal cells requires the GW182 protein. GW182 is recruited to the miRNA repression complex via interaction with Argonaute protein, and functions downstream to repress protein synthesis. Interaction with Argonaute is mediated by GW/WG repeats, whic...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq501
更新日期:2010-10-01 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::Aspects of the metabolism of RNA have been studied in enzymatically isolated protoplasts from cotyledon and first leaf mesophyll tissue of two cultivars of cucumber. The first leaf mesophyll protoplasts incorporated (3H)-uridine into ribosomal RNA at a constant rate for up to 25 hr in a simple salts medium and for up ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/2.7.1111
更新日期:1975-07-01 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
abstract::Alternative splicing is the primary source of proteome complexity in metazoans and its regulation shapes the proteome in response to shifting physiological requirements. We developed a bichromatic splicing reporter that uses a peculiar feature of some fluorescent protein coding regions to express two different fluores...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkl967
更新日期:2006-01-01 00:00:00
abstract::The 'druggable genome' encompasses several protein families, but only a subset of targets within them have attracted significant research attention and thus have information about them publicly available. The Illuminating the Druggable Genome (IDG) program was initiated in 2014, has the goal of developing experimental...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw1072
更新日期:2017-01-04 00:00:00
abstract::A novel expression system was established that allows expression and propagation of the human hepatitis B virus (HBV) genome in cultured animal cells. An overlength HBV genome encoding the viral pregenomic RNA was put under transcriptional control of the human metallothionein IIA promoter thereby replacing the endogen...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/15.24.10117
更新日期:1987-12-23 00:00:00
abstract::As the largest group of MYB family transcription factors, R2R3-MYB proteins play essential roles during plant growth and development. However, the structural basis underlying how R2R3-MYBs recognize the target DNA remains elusive. Here, we report the crystal structure of Arabidopsis WEREWOLF (WER), an R2R3-MYB protein...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkz1081
更新日期:2020-01-10 00:00:00
abstract::The unknown modified nucleotide G*, isolated from both Schizosaccharomyces pombe and Torulopsis utilis initiator tRNAs(Met), has been identified as an O-ribosyl-(1"----2')-guanosine-5"-phosphate, called Gr(p), by means of HPLC, UV-absorption, mass spectrometry and periodate oxidation procedures. By comparison with the...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/19.19.5199
更新日期:1991-10-11 00:00:00
abstract::Gene expression in Plasmodium falciparum is tightly regulated to ensure successful propagation of the parasite throughout its complex life cycle. The earliest transcriptomics studies in P. falciparum suggested a cascade of transcriptional activity over the course of the 48-hour intraerythrocytic developmental cycle (I...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx464
更新日期:2017-07-27 00:00:00
abstract::OGG1 initiated base excision repair (BER) is the major pathway for repair of oxidative DNA base damage 8-oxoguanine (8-oxoG). Here, we report that RECQL4 DNA helicase, deficient in the cancer-prone and premature aging Rothmund-Thomson syndrome, physically and functionally interacts with OGG1. RECQL4 promotes catalytic...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa392
更新日期:2020-07-09 00:00:00
abstract::The genome-wide identification of microRNA transcription start sites (miRNA TSSs) is essential for understanding how miRNAs are regulated in development and disease. In this study, we developed mirSTP (mirna transcription Start sites Tracking Program), a probabilistic model for identifying active miRNA TSSs from nasce...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx318
更新日期:2017-07-27 00:00:00
abstract::Proper DNA double-strand break (DSB) repair is essential for maintaining genome integrity. Microhomology-mediated end joining (MMEJ) is an error-prone repair mechanism, which introduces mutations at break sites and contributes to chromosomal translocations and telomere fusions, thus driving carcinogenesis. Mitotic kin...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky810
更新日期:2018-11-16 00:00:00
abstract::In Saccharomyces cerevisiae, RNA polymerase II assembly is probably initiated by the formation of the RPB3-RPB11 heterodimer. RPB3 is encoded by a single copy gene in the yeast, mouse and human genomes. The RPB11 gene is also unique in yeast and mouse, but in humans a gene family has been identified that potentially e...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gki672
更新日期:2005-06-24 00:00:00
abstract::Adaptation of the P1 phage-derived Cre /loxP site- specific recombination system to the gene targeting technique allows for the conditional deletion of genes in mice. To selectively modify genes in B lymphocytes, we have generated mice (designated CD19-Cre) which express cre under the transcriptional control of the B ...
journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:1997-03-15 00:00:00
abstract::The promoter of the human BCR gene, regulating the transcription of the chimeric BCR/ABL mRNA in leukemia, has been isolated and characterized. A region of 1.1 kb immediately 5' to the transcription start site was analyzed in detail by sequencing, DNase 1 footprinting, gel retardation and functional studies. These exp...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.23.7119
更新日期:1990-12-11 00:00:00
abstract::Type I toxin-antitoxin (TA) systems have been identified in a wide range of bacterial genomes. Here, we report the characterization of a new type I TA system present on the chromosome of the major human gastric pathogen, Helicobacter pylori. We show that the aapA1 gene encodes a 30 amino acid peptide whose artificial ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw1343
更新日期:2017-05-05 00:00:00
abstract::The effect of amino groups attached at positions 2 and 8 of the hypoxanthine moiety in the structure, reactivity and stability of DNA duplexes and triplexes is studied by means of quantum mechanical calculations, as well as extended molecular dynamics (MD) and thermodynamic integration (MD/TI) simulations. Theoretical...
journal_title:Nucleic acids research
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doi:10.1093/nar/29.12.2522
更新日期:2001-06-15 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 sequenced the 480 base pair (bp) repeating unit of the 5S RNA genes of the Dipteran fly Calliphora erythrocephala and compared this sequence to the three known 5S RNA gene sequences from the Dipteran Genus Drosophila (1,2). A striking series of five perfectly conserved homologies identically positioned within ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/12.21.8193
更新日期:1984-11-12 00:00:00
abstract::Recent genome-wide analyses have elucidated the extent of alternative splicing (AS) in mammals, often focusing on comparisons of splice isoforms between differentiated tissues. However, regulated splicing changes are likely to be important in biological transitions such as cellular differentiation, or response to envi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq614
更新日期:2010-11-01 00:00:00
abstract::To successfully infect plants, viruses must counteract small RNA-based host defense responses. During infection of Arabidopsis, Cauliflower mosaic pararetrovirus (CaMV) is transcribed into pregenomic 35S and subgenomic 19S RNAs. The 35S RNA is both reverse transcribed and also used as an mRNA with highly structured 60...
journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:2011-07-01 00:00:00
abstract::The mode of action of regulated promoters is largely determined by kinetic parameters which govern the interaction between promoters and proteins involved in induction and repression of transcription. To gain insight into the interplay between positively and negatively acting transcriptional regulators, in this case A...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/29.18.3873
更新日期:2001-09-15 00:00:00
abstract::The HIV-1 nucleocapsid (NC) protein is a small, basic protein containing two retroviral zinc fingers. It is a highly active nucleic acid chaperone; because of this activity, it plays a crucial role in virus replication as a cofactor during reverse transcription, and is probably important in other steps of the replicat...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkj442
更新日期:2006-01-24 00:00:00