Abstract:
:The use of bacterial artificial chromosomes (BACs) provides a consistent and high targeting efficiency of homologous recombination in embryonic stem (ES) cells, facilitated by long stretches of sequence homology. Here, we introduce a BAC targeting method which employs restriction fragment length polymorphisms (RFLPs) in targeted polymorphic C57BL/6/Cast/Ei F1 mouse ES cell lines to identify properly targeted ES cell clones. We demonstrate that knockout alleles can be generated either by targeting of an RFLP located in the open reading frame thereby disrupting the RFLP and ablating gene function, or by introduction of a transcription stop cassette that prematurely stops transcription of an RFLP located downstream of the stop cassette. With both methods we have generated Rnf12 heterozygous knockout ES cells, which were identified by allele specific PCR using genomic DNA or cDNA as a template. Our results indicate that this novel strategy is efficient and precise, by combining a high targeting efficiency with a convenient PCR based readout and reliable detection of correct targeting events.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Barakat TS,Rentmeester E,Sleutels F,Grootegoed JA,Gribnau Jdoi
10.1093/nar/gkr550subject
Has Abstractpub_date
2011-10-01 00:00:00pages
e121issue
18eissn
0305-1048issn
1362-4962pii
gkr550journal_volume
39pub_type
杂志文章abstract::Eukaryotic tRNAs are transcribed as precursors. A 5'-end leader and 3'-end trailer are endonucleolytically removed by RNase P and 3'-tRNase before 3'-end CCA addition, aminoacylation, nuclear export and translation. 3'-End -CC can be a 3'-tRNase anti-determinant with the ability to prevent mature tRNA from recycling t...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/29.21.4334
更新日期:2001-11-01 00:00:00
abstract::DNA assembly allows individual DNA constructs or libraries to be assembled quickly and reliably. Most methods are either: (i) Modular, easily scalable and suitable for combinatorial assembly, but leave undesirable 'scar' sequences; or (ii) bespoke (non-modular), scarless but less suitable for construction of combinato...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky1182
更新日期:2019-02-20 00:00:00
abstract::Histone proteins are synthesized in large amounts during S-phase to package the newly replicated DNA, and are among the most stable proteins in the cell. The replication-dependent (RD)-histone mRNAs expressed during S-phase end in a conserved stem-loop rather than a polyA tail. In addition, there are replication-indep...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw620
更新日期:2016-11-02 00:00:00
abstract::Extensive efforts have been directed at the discovery, investigation and clinical monitoring of targeted therapeutics. These efforts may be facilitated by the convenient access of the genetic, proteomic, interactive and other aspects of the therapeutic targets. Here, we describe an update of the Therapeutic target dat...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx1076
更新日期:2018-01-04 00:00:00
abstract::We recently developed a new type of pyrrole (Py)-imidazole (Im) polyamide-tetrahydrocyclopropabenzindolone (CBI) conjugate with an indole linker as a stable sequence-specific alkylating agent. In this study, we investigated the gene silencing activities of polyamides A, B and C, which selectively alkylate specific seq...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkl005
更新日期:2006-02-25 00:00:00
abstract::One of the greatest challenges that modern molecular biology is facing is the understanding of the complex mechanisms regulating gene expression. A fundamental step in this process requires the characterization of regulatory motifs playing key roles in the regulation of gene expression at transcriptional and post-tran...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh465
更新日期:2004-07-01 00:00:00
abstract::Topoisomerase II is an essential enzyme that is required for virtually every process that requires movement of DNA within the nucleus or the opening of the double helix. This enzyme helps to regulate DNA under- and overwinding and removes knots and tangles from the genetic material. In order to carry out its critical ...
journal_title:Nucleic acids research
pub_type: 杂志文章,评审
doi:10.1093/nar/gkn937
更新日期:2009-02-01 00:00:00
abstract::PROtein Domain Organization and Comparison (PRODOC) comprises several programs that enable convenient comparison of proteins as a sequence of domains. The in-built dataset currently consists of approximately 698 000 proteins from 192 organisms with complete genomic data, and all the SWISSPROT proteins obtained from th...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gki474
更新日期:2005-07-01 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::Microbial enzymes have many known applications as biocatalysts in biotechnology, agriculture, medical and other industries. However, only a few enzymes are currently employed for such commercial applications. In this scenario, the current onslaught of metagenomic data provides a new unexplored treasure trove of genomi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkp1001
更新日期:2010-01-01 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::We have determined the three-dimensional structure of a non-selfcomplementary nonanucleotide duplex which contains an abasic (apyrimidinic) site in the centre, i.e. a deoxyribose residue opposite an adenosine. The majority of the base and sugar proton resonances were assigned by NOESY, COSY and 2DQF spectra in D2O and...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/15.19.8003
更新日期:1987-10-12 00:00:00
abstract::Pnkp is the end-healing and end-sealing component of an RNA repair system present in diverse bacteria from many phyla. Pnkp is composed of three catalytic modules: an N-terminal polynucleotide 5' kinase, a central 2',3' phosphatase and a C-terminal ligase. The phosphatase module is a Mn(2+)-dependent phosphodiesterase...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt221
更新日期:2013-06-01 00:00:00
abstract::Peptidome similarity analysis enables researchers to gain insights into differential peptide profiles, providing a robust tool to discriminate strain-specific peptides, true intra-species differences among biological replicates or even microorganism-phenotype variations. However, no in silico peptide fingerprinting so...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx389
更新日期:2017-07-03 00:00:00
abstract::Our web server, PIZSA (http://cospi.iiserpune.ac.in/pizsa), assesses the likelihood of protein-protein interactions by assigning a Z Score computed from interface residue contacts. Our score takes into account the optimal number of atoms that mediate the interaction between pairs of residues and whether these contacts...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkz368
更新日期:2019-07-02 00:00:00
abstract::The bacteriophage Mu A and B genes, which lie adjacent to each other and are colinear on the phage genome, encode proteins that catalyze efficient transposition of Mu DNA. We show that the molar ratio of A and B proteins is approximately 1:20 in extracts prepared after induction of cells containing a Mu lysogen or a p...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/16.23.11285
更新日期:1988-12-09 00:00:00
abstract::The stability of chromosome ends, the telomeres, is dependent on the ribonucleoprotein telomerase. In vitro, telomerase requires at least one RNA molecule and a reverse transcriptase-like protein. However, for telomere homeostasis in vivo, additional proteins are required. Telomerase RNAs of different species vary in ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt514
更新日期:2013-09-01 00:00:00
abstract::Mitochondrial DNA (mtDNA) resides in a high ROS environment and suffers more mutations than its nuclear counterpart. Increasing evidence suggests that mtDNA mutations are not the results of direct oxidative damage, rather are caused, at least in part, by DNA replication errors. To understand how the mtDNA replicase, P...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkz1018
更新日期:2020-01-24 00:00:00
abstract::The Mouse Genome Database (MGD) forms the core of the Mouse Genome Informatics (MGI) system (http://www.informatics.jax.org), a model organism database resource for the laboratory mouse. MGD provides essential integration of experimental knowledge for the mouse system with information annotated from both literature an...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gki113
更新日期:2005-01-01 00:00:00
abstract::Posttranslational modifications (PTMs) of histones represent a crucial regulatory mechanism of nucleosome and chromatin dynamics in various of DNA-based cellular processes, such as replication, transcription and DNA damage repair. Lysine succinylation (Ksucc) is a newly identified histone PTM, but its regulation and f...
journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:2020-09-25 00:00:00
abstract::Annotation of herpesvirus genomes has traditionally been undertaken through the detection of open reading frames and other genomic motifs, supplemented with sequencing of individual cDNAs. Second generation sequencing and high-density microarray studies have revealed vastly greater herpesvirus transcriptome complexity...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw629
更新日期:2016-10-14 00:00:00
abstract::The nucleotide sequences 3' to the translational termination codons in a collection of human genes have been analysed for evidence of a preferred 3' context for natural UAG codons. The aim was to see whether human UAG contexts can be related to the recent demonstration of the effects of 3' context on nonsense suppress...
journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:1994-01-11 00:00:00
abstract::The herpes simplex virus protein VP16 interacts with cellular factors, including the protein Oct-1, to activate viral immediate early (IE) gene transcription. We have reproduced this effect by addition of purified, full-length VP16 and the DNA-binding 'POU' domain of Oct-1 (Oct-1/POU) to a HeLa cell in vitro transcrip...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/21.24.5570
更新日期:1993-12-11 00:00:00
abstract::Reactome (http://www.reactome.org) is a manually curated open-source open-data resource of human pathways and reactions. The current version 46 describes 7088 human proteins (34% of the predicted human proteome), participating in 6744 reactions based on data extracted from 15 107 research publications with PubMed link...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt1102
更新日期:2014-01-01 00:00:00
abstract::The catalytic function of the hairpin ribozyme has been investigated by modification-interference analysis of both ribozyme and substrate, using ribonucleoside phosphorothioates. Thiophosphate substitutions in two ribozyme domains were examined by using a novel and highly efficient two-piece ribozyme assembled from tw...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/20.11.2835
更新日期:1992-06-11 00:00:00
abstract::Cisplatin analogues were synthesised that consisted of platinum(II) diamine complexes tethered via a polymethylene chain ( n = 3, 5, 8 and 10) to a phenanthridinium cation. Both chloro and iodo leaving groups were examined. DNA adduct formation was quantitatively analysed using a linear amplification system with the p...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.17.3933
更新日期:1998-09-01 00:00:00
abstract::The aminoacylation kinetics of T7 transcripts representing defined regions of Escherichia coli serine tRNAs were determined using purified E.coli seryl-tRNA synthetase (SerRS) and the kinetic values were used to estimate the relative contribution of various tRNA(Ser) domains to recognition by SerRS. The analysis revea...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/21.19.4467
更新日期:1993-09-25 00:00:00
abstract::Infrared and UV spectroscopies have been used to study the assembly of a hairpin nucleotide sequence (nucleotides 3-30) of the 5' non-coding region of the hepatitis C virus RNA (5'-GGCGGGGAUUAUCCCCGCUGUGAGGCGG-3') with a RNA 20mer ligand (5'-CCGCCUCACAAAGGUGGGGU-3') in the presence of magnesium ion and spermidine. The...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/30.6.1333
更新日期:2002-03-15 00:00:00
abstract::Gel migration and uranyl photoprobing have been used to study the effects of inosine and 2,6-diaminopurine (2,6-DAP) substitution on adenine-tract (A-tract) induced DNA curvature. Using a 10mer repeated sequence including five inosines we show by uranyl photoprobing that a narrow minor groove varying in phase with the...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/25.17.3497
更新日期:1997-09-01 00:00:00
abstract::The polypeptides recognized by a monoclonal antibody against calf thymus DNA polymerase alpha (secreted from a hybridoma CL22 -2- 42B , Nucleic Acids Res. (1982) 10, 4703-4713) were identified by the immunoblot method as the large polypeptides of the partially-purified 10S DNA polymerase alpha fraction. Using an immun...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/12.11.4455
更新日期:1984-06-11 00:00:00