Abstract：
:Lambda-red system-based recombinogenic engineering is a powerful new method to engineer DNA without the need for restriction enzymes or ligases. Here, we report the use of a single selectable marker to enhance the usefulness of this approach. The strategy is to utilize the thymidylate synthase A (thyA) gene, which encodes an enzyme involved in the synthesis of thymidine 5'-triphosphate, for both positive and negative selection. With this approach, we successfully created point mutations in plasmid and bacterial artificial chromosome (BAC) DNA containing the mouse Col10a1 gene. The results showed that the thyA selection system is highly efficient and accurate, giving an average of >90% selection efficiency. This selection system produces DNA that is free from permanent integration of unwanted sequences, thus allowing unlimited rounds of modifications if required.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Wong QN,Ng VC,Lin MC,Kung HF,Chan D,Huang JDdoi
10.1093/nar/gni059keywords:
subject
Has Abstractpub_date
2005-03-30 00:00:00pages
e59issue
6eissn
0305-1048issn
1362-4962pii
33/6/e59journal_volume
33pub_type
杂志文章abstract：:The DNA damage checkpoint halts cell cycle progression in G2 in response to genotoxic insults. Central to the execution of cell cycle arrest is the checkpoint-induced stabilization of securin-separase complex (yeast Pds1-Esp1). The checkpoint kinases Chk1 and Chk2 (yeast Chk1 and Rad53) are thought to critically contr...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkaa355
更新日期：2020-06-19 00:00:00
abstract：:In plants, nuclear multisubunit RNA polymerases IV and V are RNA Polymerase II-related enzymes that synthesize non-coding RNAs for RNA-directed DNA methylation (RdDM) and transcriptional gene silencing. Here, we tested the importance of the C-terminal domain (CTD) of Pol IV's largest subunit given that the Pol II CTD ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkz615
更新日期：2019-09-26 00:00:00
abstract：:E. coli C6 rel- met- cys- was cultured in a fully supplemented medium and in media lacking cysteine or methionine. tRNA isolated from the three cultures containted, respectively, a normal complement of modified nucleosides; a deficiency in thiolated nucleosides and a deficiency in methylated nucleosides. Both sulfur-d...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/2.5.691
更新日期：1975-05-01 00:00:00
abstract：:A list of DNA sequences cloned from the human genome is presented. Intended as a guide to clone availability, this list includes published reports of cDNA, genomic and synthetic clones comprising gene and pseudogene sequences, uncharacterised DNA segments and repetitive DNA elements. ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/16.suppl.r403
更新日期：1988-01-01 00:00:00
abstract：:The Pk-rec gene, encoding a RecA/RAD51 homologue from the hyperthermophilic archaeon Pyrococcussp. KOD1, was expressed in Escherichia coli. The recombinant Pk-REC was purified to homogeneity and was shown to be in a dimeric form. A striking property of the purified recombinant Pk-REC was the unusual DNase activity on ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/25.4.719
更新日期：1997-02-15 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：:To investigate the feasibility of conducting a genomic-scale protein labeling and localization study in Escherichia coli, a representative subset of 23 coding DNA sequences (CDSs) was selected for chromosomal tagging with one or more fluorescent protein genes (EGFP, EYFP, mRFP1, DsRed2). We used lambda-Red recombinati...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkl1158
更新日期：2007-01-01 00:00:00
abstract：:The HSV gene encoding ICP4 is negatively regulated and the HSV gene encoding thymidine kinase is positively regulated by ICP4 in vivo. We report that ICP4 is a component of a stable complex that contains protein and a sequence of approximately 28 nucleotides that span the ICP4 gene transcription initiation site. The a...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/16.2.555
更新日期：1988-01-25 00:00:00
abstract：:A refined map for the linear arrangement of histones along DNA in nucleosomal core particles has been determined by DNA-protein crosslinking. On one strand of 145-bp core DNA, histones are aligned in the following order: (5') H2B25,35-H455,65-H375,85,95/H488-H2B105,11 5-H2A118-H3135,145/H2A145 (3') (the subscripts giv...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/13.10.3439
更新日期：1985-05-24 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：:Bacteria adapt to changes in their environment via differential gene expression mediated by DNA binding transcriptional regulators. The PRODORIC2 database hosts one of the largest collections of DNA binding sites for prokaryotic transcription factors. It is the result of the thoroughly redesigned PRODORIC database. PR...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkx1091
更新日期：2018-01-04 00:00:00
abstract：:Observation and tracking of fluorescently labeled molecules and particles in living cells reveals detailed information about intracellular processes on the molecular level. Whereas light microscopic particle observation is usually limited to two-dimensional projections of short trajectory segments, we report here imag...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gku1194
更新日期：2015-01-01 00:00:00
abstract：:PCAF and hGCN5 are distinct human genes that encode proteins related to the yeast histone acetyltransferase and transcriptional adapter GCN5. The PCAF protein shares extensive similarity with the 439 amino acids of yGCN5, but it has an approximately 350 amino acid N-terminal extension that interacts with the transcrip...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/26.12.2948
更新日期：1998-06-15 00:00:00
abstract：:In this article, we introduce the Gypsy Database (GyDB) of mobile genetic elements, an in-progress database devoted to the non-redundant analysis and evolutionary-based classification of mobile genetic elements. In this first version, we contemplate eukaryotic Ty3/Gypsy and Retroviridae long terminal repeats (LTR) ret...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkm697
更新日期：2008-01-01 00:00:00
abstract：:The side chains of the 20 types of amino acids, owing to a large extent to their different physical properties, have characteristic distributions in interior/surface regions of individual proteins and in interface/non-interface portions of protein surfaces that bind proteins or nucleic acids. These distributions have ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkm231
更新日期：2007-07-01 00:00:00
abstract：:The Biodegradative Strain Database (BSD) is a freely-accessible, web-based database providing detailed information on degradative bacteria and the hazardous substances that they degrade, including corresponding literature citations, relevant patents and links to additional web-based biological and chemical data. The B...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkg032
更新日期：2003-01-01 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：:The entire intervening sequence of Tetrahymena thermophila ribosomal DNA has been determined. It is 413 nucleotides long and has the same splice junctions as those in T. pigmentosa. There is 93% homology between the intervening sequences in the two species, and 100% homology between their adjacent 26S RNA coding regio...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/10.9.2809
更新日期：1982-05-11 00:00:00
abstract：:The distribution and amount of 5-methylcytosine (5-MeCyt) in DNA was measured for early embryos of mouse strain CF1 (2 to 4 cell stage to blastocyst) and mouse teratocarcinoma cells. In each case, the pattern of methylation was examined by use of the restriction enzymes Hha I and HPA II HPA II, which cut DNA at the si...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/7.8.2369
更新日期：1979-12-20 00:00:00
abstract：:Biological evolution resulted in a homochiral world in which nucleic acids consist exclusively of d-nucleotides and proteins made by ribosomal translation of l-amino acids. From the perspective of synthetic biology, however, particularly anabolic enzymes that could build the mirror-image counterparts of biological mac...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkx079
更新日期：2017-04-20 00:00:00
abstract：:Ribosome biogenesis in Saccharomyces cerevisiae involves a regulon of >200 genes (Ribi genes) coordinately regulated in response to nutrient availability and cellular growth rate. Two cis-acting elements called PAC and RRPE are known to mediate Ribi gene repression in response to nutritional downshift. Here, we show t...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkx058
更新日期：2017-05-05 00:00:00
abstract：:Photoalkylation reactions with 2-propanol, initiated with di-tert-butyl peroxide, of a variety of purine and pyrimidine mononucleotides and dinucleoside monophosphates lead to the substitution of an alpha-hydroxyisopropyl group for the H-8 atom of adenosine and the addition of the alcohol across the 5,6-double bond of...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/3.7.1715
更新日期：1976-07-01 00:00:00
abstract：:In neural stem cells (NSCs), the balance between stem cell maintenance and neuronal differentiation depends on cell-fate determinants such as TRIM32. Previously, we have shown that TRIM32 associates with the RNA-induced silencing complex and increases the activity of microRNAs such as Let-7a. However, the exact mechan...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkv138
更新日期：2015-03-11 00:00:00
abstract：:Rad51 protein promotes homologous recombination in eukaryotes. Recombination activities are activated by Rad51 filament assembly on ssDNA. Previous studies of yeast Rad51 showed that His352 occupies an important position at the filament interface, where it could relay signals between subunits and active sites. To inve...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gkq209
更新日期：2010-08-01 00:00:00
abstract：:The human gene for plasminogen activator inhibitor type-1 (PAI-1) has been isolated and its promoter region characterized. PAI-1 regulation by glucocorticoids, transforming growth factor-beta (TGF-beta) and the phorbol ester PMA is shown to be exerted at the promoter level. A fragment spanning 805 nucleotides of the 5...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/16.7.2805
更新日期：1988-04-11 00:00:00
abstract：:By putting heterologous genomic regulatory systems into contact, chromosome addition lines derived from interspecific or intergeneric crosses allow the investigation of transcriptional regulation in new genomic environments. Here, we report the transcriptional and epigenetic adaptation of stably inherited alien maize ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/gky209
更新日期：2018-06-01 00:00:00
abstract：:The secondary structure of an adenovirus associated low molecular weight RNA (VAI-RNA) has been studied by partial digestion with T1-RNase and S1-endonuclease followed by T1-fingerprint analysis. The empirical secondary structure has been compared with two computer generated models based on minimal free energy of the ...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/9.17.4239
更新日期：1981-09-11 00:00:00
abstract：:We have assessed the ability of nucleosomes to influence the formation of mammalian topoisomerase II-DNA complexes by mapping the sites of cleavage induced by four unrelated topoisomerase II inhibitors in naked versus nucleosome-reconstituted SV40 DNA. DNA fragments were reconstituted with histone octamers from HeLa c...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/18.15.4553
更新日期：1990-08-11 00:00:00
abstract：:A cross index is presented for using the improved selectivity offered by the Hung and Wensink (Nucl. Acids Res. 12, 1863-1874, 1984) method of partially filling in 5'-extensions produced by type II restriction endonucleases. After this treatment, DNA fragments which normally cannot be ligated to one another, can be jo...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/15.8.3199
更新日期：1987-04-24 00:00:00
abstract：:Improving the accuracy of prediction of gene starts is one of a few remaining open problems in computer prediction of prokaryotic genes. Its difficulty is caused by the absence of relatively strong sequence patterns identifying true translation initiation sites. In the current paper we show that the accuracy of gene s...
journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/29.12.2607
更新日期：2001-06-15 00:00:00
