Abstract:
:In vitro compartmentalization (IVC) was employed for the first time to select for novel bacteriophage lambda integrase variants displaying significantly enhanced recombination activity on a non-cognate target DNA sequence. These variants displayed up to 9-fold increased recombination activity over the parental enzyme, and one mutant recombined the chosen non-cognate substrate more efficiently than the parental enzyme recombined the wild-type DNA substrate. The in vitro specificity phenotype extended to the intracellular recombination of episomal vectors in HEK293 cells. Surprisingly, mutations conferring the strongest phenotype do not occur in the lambda integrase core-binding domain, which is known to interact directly with cognate target sequences. Instead, they locate to the N-terminal domain which allosterically modulates integrase activity, highlighting a previously unknown role for this domain in directing integrase specificity. The method we describe provides a robust, completely in vitro platform for the development of novel integrase reagent tools for in vitro DNA manipulation and other biotechnological applications.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Tay Y,Ho C,Droge P,Ghadessy FJdoi
10.1093/nar/gkp1089subject
Has Abstractpub_date
2010-03-01 00:00:00pages
e25issue
4eissn
0305-1048issn
1362-4962pii
gkp1089journal_volume
38pub_type
杂志文章abstract::Bacterial RNase III plays important roles in the processing and degradation of RNA transcripts. A major goal is to identify the cleavage targets of this endoribonuclease at a transcriptome-wide scale and delineate its in vivo cleavage rules. Here we applied to Escherichia coli grown to either exponential or stationary...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky684
更新日期:2018-11-02 00:00:00
abstract::In the haploid genome of Xenopus laevis there are two genes coding for the r-protein L14. It is not known if they are located on the same chromosome. cDNA clones deriving from the transcripts of the two genes have been isolated from an oocyte messenger cDNA bank showing that they are both expressed. We have studied th...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/14.19.7633
更新日期:1986-10-10 00:00:00
abstract::An updated list of DNA sequences cloned from the human genome over the past year (1991) is presented. Intended as a guide to clone availability, this list includes published reports of cDNA, genomic and synthetic probes comprising both gene and pseudogene sequences. ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/20.suppl.2181
更新日期:1992-05-11 00:00:00
abstract::The two-step process of selection and counter-selection is a standard way to enable genetic modification and engineering of bacterial genomes using homologous recombination methods. The tetA and sacB genes are contained in a DNA cassette and confer a novel dual counter-selection system. Expression of tetA confers bact...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt1075
更新日期:2013-12-01 00:00:00
abstract::The TGGCA protein, the chicken homologue of HeLa cell NF-I, was purified to homogeneity from liver tissue by a procedure which includes preparative mobility shift electrophoresis (PMSE) as the final step. PMSE was here adjusted for the isolation of the TGGCA protein, but can be used as a general method to characterize...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/15.23.9707
更新日期:1987-12-10 00:00:00
abstract::The Protein Circular Dichroism Data Bank (PCDDB) has been in operation for more than 5 years as a public repository for archiving circular dichroism spectroscopic data and associated bioinformatics and experimental metadata. Since its inception, many improvements and new developments have been made in data display, se...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw796
更新日期:2017-01-04 00:00:00
abstract::Gene trap vectors developed for genome-wide mutagenesis can be used to study factors governing the expression of exons inserted throughout the genome. For example, entrapment vectors consisting of a partial 3'-terminal exon [i.e. a neomycin resistance gene (Neo), a poly(A) site, but no 3' splice site] were typically e...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh604
更新日期:2004-05-20 00:00:00
abstract::RNA thermometers control the translation of several heat shock and virulence genes by their temperature-sensitive structural transitions. Changes in the structure and dynamics of MiniROSE RNA, which regulates translation in the temperature range of 20-45°C, were studied by site specifically replacing seven adenine res...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gku1264
更新日期:2015-01-01 00:00:00
abstract::We have examined the ability of a cationic planar chromophore, ethidium bromide, to intercalate into a short, defined triple helix. Using UV absorption, fluorescence spectroscopy and a gel retardation assay we demonstrate that ethidium bromide is able to bind to a triple helix with a lower affinity than to the corresp...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/19.7.1521
更新日期:1991-04-11 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::Frequencies for each of the 61 amino acid codons have been determined in every published mRNA sequence of 50 or more codons. The frequencies are shown for each kind of genome and for each individual gene. A surprising consistency of choices exists among genes of the same or similar genomes. Thus each genome, or kind o...
journal_title:Nucleic acids research
pub_type: 杂志文章,评审
doi:10.1093/nar/8.1.197-c
更新日期:1980-01-11 00:00:00
abstract::The IMGT/HLA database (http://www.ebi.ac.uk/imgt/hla) has provided a centralized repository for the sequences of the alleles named by the WHO Nomenclature Committee for Factors of the HLA System for the past four years. Since its initial release the database has grown and is the primary source of information for the s...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkg070
更新日期:2003-01-01 00:00:00
abstract::Influenza A viruses are hosted by numerous avian and mammalian species, which have shaped their evolution into distinct lineages worldwide. The viral genome consists of eight RNA segments that are frequently exchanged between different viruses via a process known as genetic reassortment. A complete genotype nomenclatu...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkm365
更新日期:2007-07-01 00:00:00
abstract::RNA:5-methylcytosine (m5C) methyltransferases are currently the focus of intense research following a series of high-profile reports documenting their physiological links to several diseases. However, no methods exist which permit the specific analysis of RNA:m5C methyltransferases in cells. Herein, we described how a...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkz1047
更新日期:2020-01-10 00:00:00
abstract::Previous work in predicting protein localization to the chloroplast organelle in plants led to the development of an artificial neural network-based approach capable of remarkable accuracy in its prediction (ChloroP). A common criticism against such neural network models is that it is difficult to interpret the criter...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/29.16.e82
更新日期:2001-08-15 00:00:00
abstract::The reported binding preference of human hnRNP protein A1 for the 3'-splice site of some introns (Swanson and Dreyfuss (1988) EMBO J. 7, 3519-3529; Mayrand and Pederson (1990) Nucleic Acids Res. 18, 3307-3318) was tested by assaying in vitro the binding of purified recombinant A1 protein (expressed in bacteria) to syn...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.22.6595
更新日期:1990-11-25 00:00:00
abstract::Data produced with short-read sequencing technologies result in ambiguous haplotyping and a limited capacity to investigate the full repertoire of biologically relevant forms of genetic variation. The notion of haplotype-resolved sequencing data has recently gained traction to reduce this unwanted ambiguity and enable...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx436
更新日期:2017-07-27 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::A newly discovered group of spherical plant viruses contains a bipartite genome consisting of a single-strand linear RNA molecule (RNA 1, Mr 1.5 x 10(6) ), and a single-strand, covalently closed circular viroid-like RNA molecule (RNA 2, Mr approximately 125,000). The nucleotide sequences of the RNA 2 of two of these, ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/10.12.3681
更新日期:1982-06-25 00:00:00
abstract::The hexanucleoside pentaphosphate d(m5CpGpm5CpGpm5CpG) has been studied in solution by ultra-violet absorption, circular dichroism and 31P nuclear magnetic resonance under various experimental conditions. In 0.2 M NaClO4 at low temperature, an hexamer duplex is formed which has a B or B-like conformation. As the salt ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/11.13.4453
更新日期:1983-07-11 00:00:00
abstract::Telomeres comprise the protective caps of natural chromosome ends and function in the suppression of DNA damage signaling and cellular senescence. Therefore, techniques used to determine telomere length are important in a number of studies, ranging from those investigating telomeric structure to effects on human disea...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt1315
更新日期:2014-02-01 00:00:00
abstract::Extrachromosomal circular DNA (eccDNA) is both a driver of eukaryotic genome instability and a product of programmed genome rearrangements, but its extent had not been surveyed in Oxytricha, a ciliate with elaborate DNA elimination and translocation during development. Here, we captured rearrangement-specific circular...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkz725
更新日期:2019-10-10 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::Oriented DNA films prepared by the wet-spinning technique have been complexed with several ligands: the anthracycline antibiotic violamycin BI, the dipeptide L-carnosine, and the oligopeptide antibiotic netropsin. The formation of the DNA-ligand complexes is accompanied by dramatic changes of the conformational flexib...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/12.23.9165
更新日期:1984-12-11 00:00:00
abstract::For efficient DNA cleavage, the Type III restriction endonuclease EcoP15I communicates with two inversely oriented recognition sites in an ATP-dependent process. EcoP15I consists of methylation (Mod) and restriction (Res) subunits forming a multifunctional enzyme complex able to methylate or to cleave DNA. In this stu...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr1239
更新日期:2012-04-01 00:00:00
abstract::A small library of 49 peptide-oligonucleotide conjugates were synthesized to explore the influence of various peptide side chains on the hybridization properties of the DNA. An invariant 8mer oligonucleotide was coupled to a peptide portion that contained a five residue variable region composed of the cationic amino a...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.13.3136
更新日期:1998-07-01 00:00:00
abstract::Transcriptional regulatory networks specify regulatory proteins controlling the context-specific expression levels of genes. Inference of genome-wide regulatory networks is central to understanding gene regulation, but remains an open challenge. Expression-based network inference is among the most popular methods to i...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw963
更新日期:2017-02-28 00:00:00
abstract::RNA-binding proteins (RBPs) play important roles in the post-transcriptional control of RNAs. Identifying RBP binding sites and characterizing RBP binding preferences are key steps toward understanding the basic mechanisms of the post-transcriptional gene regulation. Though numerous computational methods have been dev...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv1025
更新日期:2016-02-29 00:00:00
abstract::Anti-silencing function 1 (Asf1) and Chromatin Assembly Factor 1 (CAF-1) chaperone histones H3/H4 during the assembly of nucleosomes on newly replicated DNA. To understand the mechanism of histone H3/H4 transfer among Asf1, CAF-1 and DNA from a thermodynamic perspective, we developed and employed biophysical approache...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gks906
更新日期:2012-12-01 00:00:00
abstract::RNase J is a conserved ribonuclease that belongs to the β-CASP family of nucleases. It possesses both endo- and exo-ribonuclease activities, which play a key role in pre-rRNA maturation and mRNA decay. Here we report high-resolution crystal structures of Deinococcus radiodurans RNase J complexed with RNA or uridine 5'...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv444
更新日期:2015-06-23 00:00:00