Random DNA fragmentation with endonuclease V: application to DNA shuffling.

Abstract:

:The enzyme endonuclease V nicks uracil-containing DNA at the second or third phosphodiester bond 3' to uracil sites. I applied the enzyme to random fragmentation of DNA to revise the complex DNA shuffling protocol. The merit of using endonuclease V is that cleavage occurs at random sites and the length of the fragments can easily be adjusted by varying the concentration of dUTP in the polymerase chain reaction. Unlike the conventional method using DNase I, no partial digestion or gel separation of fragments is required. Therefore, labor is dramatically reduced and reproducibility ensured. I applied this method to recombine two truncated green fluorescent protein (GFP) genes and demonstrated successful DNA shuffling by the appearance of the fluorescent full-length GFP genes.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Miyazaki K

doi

10.1093/nar/gnf139

keywords:

subject

Has Abstract

pub_date

2002-12-15 00:00:00

pages

e139

issue

24

eissn

0305-1048

issn

1362-4962

journal_volume

30

pub_type

杂志文章
  • Genetic manipulation of an exogenous non-immunoglobulin protein by gene conversion machinery in a chicken B cell line.

    abstract::During culture, a chicken B cell line DT40 spontaneously mutates immunoglobulin (Ig) genes by gene conversion, which involves activation-induced cytidine deaminase (AID)-dependent homologous recombination of the variable (V) region gene with upstream pseudo-V genes. To explore whether this mutation mechanism can targe...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gnj013

    authors: Kanayama N,Todo K,Takahashi S,Magari M,Ohmori H

    更新日期:2006-01-18 00:00:00

  • G-quadruplex formation at the 3' end of telomere DNA inhibits its extension by telomerase, polymerase and unwinding by helicase.

    abstract::Telomere G-quadruplex is emerging as a promising anti-cancer target due to its inhibition to telomerase, an enzyme expressed in more than 85% tumors. Telomerase-mediated telomere extension and some other reactions require a free 3' telomere end in single-stranded form. G-quadruplex formation near the 3' end of telomer...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkr164

    authors: Wang Q,Liu JQ,Chen Z,Zheng KW,Chen CY,Hao YH,Tan Z

    更新日期:2011-08-01 00:00:00

  • Probing sequence-specific RNA recognition by the bacteriophage MS2 coat protein.

    abstract::We present the results of in vitro binding studies aimed at defining the key recognition elements on the MS2 RNA translational operator (TR) essential for complex formation with coat protein. We have used chemically synthesized operators carrying modified functional groups at defined nucleotide positions, which are es...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/23.13.2512

    authors: Stockley PG,Stonehouse NJ,Murray JB,Goodman ST,Talbot SJ,Adams CJ,Liljas L,Valegård K

    更新日期:1995-07-11 00:00:00

  • The G-rich auxiliary downstream element has distinct sequence and position requirements and mediates efficient 3' end pre-mRNA processing through a trans-acting factor.

    abstract::A downstream G-rich sequence (GRS), GGGGGAGGUGUGGG, has been previously shown to influence the efficiency of 3' end processing of the SV40 late polyadenylation signal. We have now defined several important parameters for GRS-mediated polyadenylation. The ability of the GRS to influence 3' end processing efficiency was...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/23.9.1625

    authors: Bagga PS,Ford LP,Chen F,Wilusz J

    更新日期:1995-05-11 00:00:00

  • The thy pol-2 intein of Thermococcus hydrothermalis is an isoschizomer of PI-TliI and PI-TfuII endonucleases.

    abstract::THY Pol-2 intein, from Thermococcus hydrothermalis, belongs to the same allelic family as TLI Pol-2 (PI-TLII), Tfu Pol-2 (PI-TFUII) and TspTY Pol-3 mini-intein, all inserted at the pol-c site of archaeal DNA polymerase genes. This new intein was cloned, expressed in Escherichia coli and purified. The intein is a speci...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/28.21.4391

    authors: Saves I,Eleaume H,Dietrich J,Masson JM

    更新日期:2000-11-01 00:00:00

  • Controlled loading of oligodeoxyribonucleotide monolayers onto unoxidized crystalline silicon; fluorescence-based determination of the surface coverage and of the hybridization efficiency; parallel imaging of the process by Atomic Force Microscopy.

    abstract::Unoxidized crystalline silicon, characterized by high purity, high homogeneity, sturdiness and an atomically flat surface, offers many advantages for the construction of electronic miniaturized biosensor arrays upon attachment of biomolecules (DNA, proteins or small organic compounds). This allows to study the inciden...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gnj034

    authors: Cattaruzza F,Cricenti A,Flamini A,Girasole M,Longo G,Prosperi T,Andreano G,Cellai L,Chirivino E

    更新日期:2006-02-28 00:00:00

  • A flexible multiple sequence alignment program.

    abstract::The 'regions' method for multisequence alignment used in the previously reported program MALIGN has been generalized to include recursive refinement so that unaligned portions between two regions at the current level of resolution can be handled with increased resolution. Additionally, there is incorporated a limiting...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/16.5.1683

    authors: Martinez HM

    更新日期:1988-03-11 00:00:00

  • Functional regulation of FEN1 nuclease and its link to cancer.

    abstract::Flap endonuclease-1 (FEN1) is a member of the Rad2 structure-specific nuclease family. FEN1 possesses FEN, 5'-exonuclease and gap-endonuclease activities. The multiple nuclease activities of FEN1 allow it to participate in numerous DNA metabolic pathways, including Okazaki fragment maturation, stalled replication fork...

    journal_title:Nucleic acids research

    pub_type: 杂志文章,评审

    doi:10.1093/nar/gkq884

    authors: Zheng L,Jia J,Finger LD,Guo Z,Zer C,Shen B

    更新日期:2011-02-01 00:00:00

  • Genome-wide prediction of minor-groove electrostatic potential enables biophysical modeling of protein-DNA binding.

    abstract::Protein-DNA binding is a fundamental component of gene regulatory processes, but it is still not completely understood how proteins recognize their target sites in the genome. Besides hydrogen bonding in the major groove (base readout), proteins recognize minor-groove geometry using positively charged amino acids (sha...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkx915

    authors: Chiu TP,Rao S,Mann RS,Honig B,Rohs R

    更新日期:2017-12-01 00:00:00

  • A new method for predicting signal sequence cleavage sites.

    abstract::A new method for identifying secretory signal sequences and for predicting the site of cleavage between a signal sequence and the mature exported protein is described. The predictive accuracy is estimated to be around 75-80% for both prokaryotic and eukaryotic proteins. ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/14.11.4683

    authors: von Heijne G

    更新日期:1986-06-11 00:00:00

  • Quantitation of RNA editing substrates, products and potential intermediates: implications for developmental regulation.

    abstract::Kinetoplast mitochondrial RNA editing is the developmentally regulated post-transcriptional process of uridine insertion and deletion in mRNAs directed by short guide RNAs (gRNAs), which creates functional mRNAs. Two mechanisms are proposed: transesterification which predicts gRNA/mRNA chimeric intermediates, and enzy...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/23.4.708

    authors: Riley GR,Myler PJ,Stuart K

    更新日期:1995-02-25 00:00:00

  • GeneNet in 2005.

    abstract::The GeneNet system is designed for collection and analysis of the data on gene and metabolic networks, signal transduction pathways and kinetic characteristics of elementary processes. In the past 2 years, the GeneNet structure was considerably improved: (i) the current version of the database is now implemented using...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gki077

    authors: Ananko EA,Podkolodny NL,Stepanenko IL,Podkolodnaya OA,Rasskazov DA,Miginsky DS,Likhoshvai VA,Ratushny AV,Podkolodnaya NN,Kolchanov NA

    更新日期:2005-01-01 00:00:00

  • Structural insight into DNA-assembled oligochromophores: crystallographic analysis of pyrene- and phenanthrene-modified DNA in complex with BpuJI endonuclease.

    abstract::The use of the DNA duplex as a supramolecular scaffold is an established approach for the assembly of chromophore aggregates. In the absence of detailed structural insight, the characterization of thus assembled oligochromophores is, today, largely based on solution-phase spectroscopy. Here, we describe the crystal st...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkw644

    authors: Probst M,Aeschimann W,Chau TT,Langenegger SM,Stocker A,Häner R

    更新日期:2016-09-06 00:00:00

  • ADAR2 A-->I editing: site selectivity and editing efficiency are separate events.

    abstract::ADAR enzymes, adenosine deaminases that act on RNA, form a family of RNA editing enzymes that convert adenosine to inosine within RNA that is completely or largely double-stranded. Site-selective A-->I editing has been detected at specific sites within a few structured pre-mRNAs of metazoans. We have analyzed the edit...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkg681

    authors: Källman AM,Sahlin M,Ohman M

    更新日期:2003-08-15 00:00:00

  • Circular dichroism studies of an oligodeoxyribonucleotide containing a hairpin loop made of a hexaethylene glycol chain: conformation and stability.

    abstract::An oligodeoxyribonucleotide, d(GCTCACAAT-X-ATTGTGAGC), where X represents a hexaethylene glycol chain, was studied using circular dichroism spectroscopy. Its conformation and conformational stability were compared to those of compounds where X was replaced by four thymines and to the duplex of same sequence without lo...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/18.21.6353

    authors: Durand M,Chevrie K,Chassignol M,Thuong NT,Maurizot JC

    更新日期:1990-11-11 00:00:00

  • Modulations of the in vitro translational efficiencies of Yellow Fever virus mRNAs: interactions between coding and noncoding regions.

    abstract::As an approach to define the structural features within the 5' noncoding region of Yellow Fever virus (YFV) that modulate mRNA translational efficiency, we have studied how minor changes in this region affect the translational capacity in vitro of the corresponding mRNAs. A cDNA sequence coding for part of the YFV str...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/17.7.2463

    authors: Ruiz-Linares A,Bouloy M,Girard M,Cahour A

    更新日期:1989-04-11 00:00:00

  • The role of acetylation in rDNA transcription.

    abstract::Treatment of NIH 3T3 cells with trichostatin A (TSA), an inhibitor of histone deacetylase (HDAC), resulted in a dose-dependent increase in transcription from a rDNA reporter and from endogenous rRNA genes. Chromatin immunoprecipitation using anti-acetyl-histone H4 antibodies demonstrated a direct effect of TSA on the ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/29.20.4114

    authors: Hirschler-Laszkiewicz I,Cavanaugh A,Hu Q,Catania J,Avantaggiati ML,Rothblum LI

    更新日期:2001-10-15 00:00:00

  • Multi-forms of human MTH1 polypeptides produced by alternative translation initiation and single nucleotide polymorphism.

    abstract::The human MTH1 gene for 8-oxo-7,8-dihydrodeoxyguanosine triphosphatase, produces seven types (types 1, 2A, 2B, 3A, 3B, 4A and 4B) of mRNAs. The B-type mRNAs with exon 2b-2c segments have three additional in-frame AUGs in their 5' regions. We report here that these transcripts produce three forms of MTH1 polypeptides (...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/27.22.4335

    authors: Oda H,Taketomi A,Maruyama R,Itoh R,Nishioka K,Yakushiji H,Suzuki T,Sekiguchi M,Nakabeppu Y

    更新日期:1999-11-15 00:00:00

  • Many trypanosome messenger RNAs share a common 5' terminal sequence.

    abstract::The mRNAs for different variant surface antigens of Trypanosoma brucei start with the same 35 nucleotides. This sequence is encoded by a separate mini-exon, located in a 1.35-kb repetitive element. We have reported that trypanosomes contain many transcripts that hybridize to mini-exon probes, even if they do not make ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/12.9.3777

    authors: De Lange T,Michels PA,Veerman HJ,Cornelissen AW,Borst P

    更新日期:1984-05-11 00:00:00

  • The salt dependence of DNA recognition by NF-kappaB p50: a detailed kinetic analysis of the effects on affinityand specificity.

    abstract::The binding kinetics of NF-kappaB p50 to the Ig-kappaB site and to a DNA duplex with no specific binding site were determined under varying conditions of potassium chloride concentration using a surface plasmonresonance biosensor. Association and dissociation rate constants were measured enabling calculation of the di...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/27.4.1063

    authors: Hart DJ,Speight RE,Cooper MA,Sutherland JD,Blackburn JM

    更新日期:1999-02-15 00:00:00

  • Chemical synthesis of a gene for somatomedin C.

    abstract::A synthetic gene for somatomedin C, a human growth factor, has been assembled by a single ligation of 23 oligodeoxyribonucleotides, which were chemically synthesized by an improved solid phase phosphotriester method. ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/13.8.2959

    authors: Sproat BS,Gait MJ

    更新日期:1985-04-25 00:00:00

  • Genomes OnLine database (GOLD) v.7: updates and new features.

    abstract::The Genomes Online Database (GOLD) (https://gold.jgi.doe.gov) is an open online resource, which maintains an up-to-date catalog of genome and metagenome projects in the context of a comprehensive list of associated metadata. Information in GOLD is organized into four levels: Study, Biosample/Organism, Sequencing Proje...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gky977

    authors: Mukherjee S,Stamatis D,Bertsch J,Ovchinnikova G,Katta HY,Mojica A,Chen IA,Kyrpides NC,Reddy T

    更新日期:2019-01-08 00:00:00

  • The use of oligodeoxynucleotide probes in chaotrope-based hybridization solutions.

    abstract::Hybridization solutions containing chaotropes may be used to modulate the thermal stability (Tm or Td) of oligodeoxynucleotide (ODN) duplexes or hybrids over a 90 degrees C range. Modulation of Td allows formulation of hybridization solutions that permit ambient temperature hybridization using most combinations of pro...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/19.19.5143

    authors: Van Ness J,Chen L

    更新日期:1991-10-11 00:00:00

  • A compact form of double-stranded RNA in solutions containing poly(ethyleneglycol).

    abstract::Molecules of single-stranded ribosomal RNA and double-stranded replicative form of phage f2 RNA (dsRNA) adopt a compact form in solutions, containing sufficiently high concentrations of salt (NaCl) and polymer (PEG). However, only in the cases of native dsRNA molecules the compact particles are characterized by a regu...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/3.6.1533

    authors: Evdokimov YM,Pyatigorskaya TL,Kadikov VA,Polyvtsev OF,Doskocil J,Koudelka J,Varshavsky YM

    更新日期:1976-06-01 00:00:00

  • High-throughput single-molecule mapping links subtelomeric variants and long-range haplotypes with specific telomeres.

    abstract::Accurate maps and DNA sequences for human subtelomere regions, along with detailed knowledge of subtelomere variation and long-range telomere-terminal haplotypes in individuals, are critical for understanding telomere function and its roles in human biology. Here, we use a highly automated whole genome mapping technol...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkx017

    authors: Young E,Pastor S,Rajagopalan R,McCaffrey J,Sibert J,Mak ACY,Kwok PY,Riethman H,Xiao M

    更新日期:2017-05-19 00:00:00

  • A hand-off of DNA between archaeal polymerases allows high-fidelity replication to resume at a discrete intermediate three bases past 8-oxoguanine.

    abstract::During DNA replication, the presence of 8-oxoguanine (8-oxoG) lesions in the template strand cause the high-fidelity (HiFi) DNA polymerase (Pol) to stall. An early response to 8-oxoG lesions involves 'on-the-fly' translesion synthesis (TLS), in which a specialized TLS Pol is recruited and replaces the stalled HiFi Pol...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkaa803

    authors: Cranford MT,Kaszubowski JD,Trakselis MA

    更新日期:2020-11-04 00:00:00

  • The histone 3'-terminal stem-loop is necessary for translation in Chinese hamster ovary cells.

    abstract::The metazoan cell cycle-regulated histone mRNAs are the only known cellular mRNAs that do not terminate in a poly(A) tall. Instead, mammalian histone mRNAs terminate in a highly conserved stem-loop structure which is required for 3'-end processing and regulates mRNA stability. The poly(A) tail not only regulates trans...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/24.10.1954

    authors: Gallie DR,Lewis NJ,Marzluff WF

    更新日期:1996-05-15 00:00:00

  • Bidirectional transfer from a 24 bp border repeat of Agrobacterium tumefaciens.

    abstract::T-region transfer from wild-type Agrobacterium strains is thought to be an orientated process, starting at the right border repeat and terminating at the left border repeat of the T-region. Here we demonstrate that a right border repeat in the inverted orientation relative to the onc-genes can also mediate transfer of...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/16.21.10225

    authors: van Haaren MJ,Sedee NJ,de Boer HA,Schilperoort RA,Hooykaas PJ

    更新日期:1988-11-11 00:00:00

  • Chicken liver TGGCA protein purified by preparative mobility shift electrophoresis (PMSE) shows a 36.8 to 29.8 kd microheterogeneity.

    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

    authors: Rupp RA,Sippel AE

    更新日期:1987-12-10 00:00:00

  • Complete sequence of the amphioxus (Branchiostoma lanceolatum) mitochondrial genome: relations to vertebrates.

    abstract::The complete nucleotide sequence of the mitochondrial DNA of the amphioxus Branchiostoma lanceolatum has been determined. This mitochondrial genome is small (15 076 bp) because of the short size of the two rRNA genes and the tRNA genes. In addition, this genome contains a very short non-coding region (57 bp) with no s...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/26.13.3279

    authors: Spruyt N,Delarbre C,Gachelin G,Laudet V

    更新日期:1998-07-01 00:00:00