Abstract:
:A method for constructing yeast artificial chromosome (YAC) libraries with large insert sizes is reported. High molecular weight human DNA was partially digested with EcoRI and cloned in the vector pYAC4. When unfractionated DNA was used, the mean YAC size was 120kb. Fractionation by pulsed-field gel electrophoresis using a 'waltzer' apparatus to remove small DNA fragments increased the mean YAC size to congruent to 220kb or congruent to 370kb depending on the fractionation conditions. Ligated DNA prepared by this method was stable at 4 degrees C and routinely yielded transformation efficiencies of greater than 700 colonies/micrograms. It should be possible to extend the method to produce even larger inserts and to use high molecular weight DNA from any source.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Anand R,Villasante A,Tyler-Smith Cdoi
10.1093/nar/17.9.3425subject
Has Abstractpub_date
1989-05-11 00:00:00pages
3425-33issue
9eissn
0305-1048issn
1362-4962journal_volume
17pub_type
杂志文章abstract::Yeast uses nucleotide excision repair (NER) and photolyase (photoreactivation) to repair cyclobutane pyrimidine dimers (CPDs) generated by ultraviolet light. In active genes, NER preferentially repairs the transcribed strand (TS). In contrast, we recently showed that photolyase preferentially repairs the non-transcrib...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/25.19.3795
更新日期:1997-10-01 00:00:00
abstract::Herpes simplex type 1 virus (HSV-1) and bacteriophage λ capsids undergo considerable structural changes during self-assembly and DNA packaging. The initial steps of viral capsid self-assembly require weak, non-covalent interactions between the capsid subunits to ensure free energy minimization and error-free assembly....
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gku634
更新日期:2014-08-01 00:00:00
abstract::The transcriptional activity of the p53 tumor suppressor protein is crucial for the regulation of cell growth, apoptosis and tumor progression. The first identified p53 relative, p73, was reported to be monoallelically expressed in normal tissues. In some tumors, loss of heterozygosity was associated with overexpressi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/28.2.513
更新日期:2000-01-15 00:00:00
abstract::Type II CRISPR-Cas9 RNA-guided nucleases are widely used for genome engineering. Type II-A SpCas9 protein from Streptococcus pyogenes is the most investigated and highly used enzyme of its class. Nevertheless, it has some drawbacks, including a relatively big size, imperfect specificity and restriction to DNA targets ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkz1225
更新日期:2020-02-28 00:00:00
abstract::The assembly of the complex that forms over the simian virus 40 origin to initiate DNA replication is not well understood. This complex is composed of the virus-coded T antigen and three cellular proteins, replication protein A (RPA), DNA polymerase alpha/primase (pol/prim) and topoisomerase I (topo I) in association ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh236
更新日期:2004-02-11 00:00:00
abstract::The ability of Deinococcus radiodurans to recover from extensive DNA damage is due in part to its ability to efficiently repair its genome, even following severe fragmentation by hundreds of double-strand breaks. The single-strand annealing pathway plays an important role early during the recovery process, making use ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt759
更新日期:2013-11-01 00:00:00
abstract::In spite of the many developments in synthetic oligonucleotide (ON) chemistry and design, invasion into double-stranded DNA (DSI) under physiological salt and pH conditions remains a challenge. In this work, we provide a new ON tool based on locked nucleic acids (LNAs), designed for strand invasion into duplex DNA (DS...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt007
更新日期:2013-03-01 00:00:00
abstract::A short histone-like fusion RNA, generated when the RNA 3' processing signal from a mouse histone H4 gene is inserted into a heterologous transcription unit, becomes correctly down-regulated in G1-arrested cells of a temperature-sensitive mouse mastocytoma cell cycle mutant (21-Tb; Stauber et al., EMBO J. 5, 3297-3303...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/16.20.9399
更新日期:1988-10-25 00:00:00
abstract::Metnase is a human protein with methylase (SET) and nuclease domains that is widely expressed, especially in proliferating tissues. Metnase promotes non-homologous end-joining (NHEJ), and knockdown causes mild hypersensitivity to ionizing radiation. Metnase also promotes plasmid and viral DNA integration, and topoisom...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq339
更新日期:2010-09-01 00:00:00
abstract::Restriction endonuclease Bse634I recognizes and cleaves the degenerate DNA sequence 5'-R/CCGGY-3' (R stands for A or G; Y for T or C, '/' indicates a cleavage position). Here, we report the crystal structures of the Bse634I R226A mutant complexed with cognate oligoduplexes containing ACCGGT and GCCGGC sites, respectiv...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gks300
更新日期:2012-08-01 00:00:00
abstract::A 2.1-kb cDNA clone representing a new member of the protease inhibitor family was isolated from a human liver cDNA library. The inhibitor, named human Leuserpin 2 (hLS2), comprises 480 amino acids and contains a leucine residue at its putative reactive center. HLS2 is about 25-28% homologous to three human members of...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/14.2.1073
更新日期:1986-01-24 00:00:00
abstract::Locked nucleic acid (LNA) is a chemically modified nucleic acid with its sugar ring locked in an RNA-like (C3'-endo) conformation. LNAs show extraordinary thermal stabilities when hybridized with DNA, RNA or LNA itself. We performed molecular dynamics simulations on five isosequential duplexes (LNA-DNA, LNA-LNA, LNA-R...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkm1182
更新日期:2008-03-01 00:00:00
abstract::The multifunctional HCV core protein consists of a hydrophilic RNA interacting D1 domain and a hydrophobic D2 domain interacting with membranes and lipid droplets. The core D1 domain was found to possess nucleic acid annealing and strand transfer properties. To further understand these chaperone properties, we investi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq094
更新日期:2010-06-01 00:00:00
abstract::Antisense oligodeoxynucleotides (ODNs) have biological activity in treating various forms of cancer. The antisense effects of two types of 20mer ODNs, phosphorothioate-modified ODNs (S-ODNs) and S-ODNs with 12 2'-O-methyl groups (Me-S-ODNs), targeted to sites 109 and 277 of bcl-2 mRNA, were compared. Both types were a...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh516
更新日期:2004-04-02 00:00:00
abstract::Mouse globin messenger ribonucleoprotein (mRNP) has been isolated from reticulocyte polysomes by affinity chromatography to oligo(T)-cellulose, using a procedure modified from that of Lindberg and Sundquist. The messenger RNA and protein moieties fo the mRNP are indistinguishable from those isolated by less rapid tech...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/2.12.2251
更新日期:1975-12-01 00:00:00
abstract::DNA methylation plays critical roles in transcriptional regulation and chromatin remodeling. Differentially methylated regions (DMRs) have important implications for development, aging and diseases. Therefore, genome-wide mapping of DMRs across various temporal and spatial methylomes is important in revealing the impa...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr053
更新日期:2011-05-01 00:00:00
abstract::Transactivation by the ETS family of transcription factors, whose members share structurally conserved DNA-binding domains, is variably sensitive to methylation of their target genes. The mechanism by which DNA methylation controls ETS proteins remains poorly understood. Uncertainly also pervades the effects of hemi-m...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw528
更新日期:2016-10-14 00:00:00
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,...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkp1089
更新日期:2010-03-01 00:00:00
abstract::The class II genes of the major histocompatibility complex (MHC) encode a family of cell surface glycoproteins that present processed antigen to the T cell receptor. Class II genes are regulated coordinately, responding to both immunologic and developmental signals. Conserved sequence elements 5' to class II genes hav...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/19.6.1243
更新日期:1991-03-25 00:00:00
abstract::GGRNA (http://GGRNA.dbcls.jp/) is a Google-like, ultrafast search engine for genes and transcripts. The web server accepts arbitrary words and phrases, such as gene names, IDs, gene descriptions, annotations of gene and even nucleotide/amino acid sequences through one simple search box, and quickly returns relevant Re...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gks448
更新日期:2012-07-01 00:00:00
abstract::Restriction-free cloning (RF-cloning) is a PCR-based technology that expands on the QuikChange™ mutagenesis process originally popularized by Stratagene in the mid-1990s, and allows the insertion of essentially any sequence into any plasmid at any location. While RF-cloning is a powerful tool for the design of custom ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gks396
更新日期:2012-07-01 00:00:00
abstract::Melting of DNA in a segment bounded at both ends by regions of greater stability during electrophoresis in denaturing gradient gels show complex properties, not accommodated with standard melting theory. Compact bands of some DNA molecules become anomalously broadened at the retardation level in a denaturing gradient,...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/23.14.2775
更新日期:1995-07-25 00:00:00
abstract::The cnjC gene from the protozoan Tetrahymena thermophila was completely sequenced. The deduced gene product was found to have significant sequence similarity to the yeast and prokaryotic RNA polymerase subunits involved with subunit assembly. Since cnjC is active only during the sexual stage (conjugation) of Tetrahyme...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.10.2953
更新日期:1990-05-25 00:00:00
abstract::Besides orthologs of other eukaryotic mismatch-repair (MMR) proteins, plants encode MSH7, a paralog of MSH6. The Arabidopsis thaliana recognition heterodimers AtMSH2*MSH6 (AtMutSalpha) and AtMSH2*MSH3 (AtMutSbeta) were previously found to bind the same subsets of mismatches as their counterparts in other eukaryotes--r...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkg780
更新日期:2003-10-15 00:00:00
abstract::To screen multiple loci in small purified samples of diploid and aneuploid cells a PCR-based technique of whole genome amplification was adapted to the study of somatic lesions. DNA samples from different numbers of flow-sorted diploid and aneuploid cells from biopsies were amplified with a degenerate 15mer primer. Al...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/23.17.3488
更新日期:1995-09-11 00:00:00
abstract::Primer Design Assistant (PDA) is a web interface primer design service combined with thermodynamic theory to evaluate the fitness of primers. It runs in a Linux-Apache-MySQL-PHP structure on a PC equipped with dual CPU (Intel Pentium III 1.4 GHz) and 512 Mb of RAM. A succinct user interface of PDA is accomplished by b...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkg560
更新日期:2003-07-01 00:00:00
abstract::To investigate molecular controls of cardiomyocyte proliferation, we utilized cardiomyocytes induced to proliferate indefinitely by SV40 large T antigen (T-ag). In the T-ag-immortalized AT-1, AT-2 and HL-1 cardiomyocytes, normal cellular proteins associating with T-ag and p53 were identified, isolated and micro-sequen...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/28.15.2882
更新日期:2000-08-01 00:00:00
abstract::Transcription factor IIIA (TFIIIA) and p43 zinc finger protein form distinct complexes with 5S ribosomal RNA in Xenopus oocytes. Additionally, TFIIIA binds the internal promoter of the 5S RNA gene and supports assembly of a transcription initiation complex. Both proteins have nine tandemly repeated zinc fingers with a...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.3.703
更新日期:1998-02-01 00:00:00
abstract::Optimization of primer sequences for polymerase chain reaction (PCR) and quantitative PCR (qPCR) and reaction conditions remains an experimental challenge. We have developed a resource, PrimerBank, which contains primers that can be used for PCR and qPCR under stringent and allele-invariant amplification conditions. A...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr1013
更新日期:2012-01-01 00:00:00
abstract::In two-colour microarrays, the ratio of signal intensities of two co-hybridized samples is used as a relative measure of gene expression. Ratio-based analysis becomes complicated and inefficient in multi-class comparisons. We therefore investigated the validity of an intensity-based analysis procedure. To this end, tw...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gnh038
更新日期:2004-02-24 00:00:00