The synthesis of blocked triplet-phosphoramidites and their use in mutagenesis.

Abstract:

:A general approach for the synthesis of oligonucleotide-triplet phosphoramidites and the synthesis of four such blocks are described. A strategy was devised to minimize the number of dimer precursors needed for synthesis of a complete set of triplet-amidite blocks encoding all 20 amino acids. Whereas synthesis of 20 triplet-amidite blocks consisting of codon sequences requires 16 dimer blocks, just seven dimer blocks are required to synthesize all required antisense sequences. The antisense sequences are then converted to codons in template mediated replication. Using a mixture of four triplet-amidites and conventional automated solid-phase DNA synthesis, short (6mer) and medium length (30mer) oligonucleotide mixtures were synthesized and analyzed. The latter was replicated in vitro and used as a mutagenic cassette to produce four mutants of Asp 221 in the enzyme thymidylate synthase. The method establishes the direction and utility for the production and use of triplet-amidite blocks in DNA synthesis.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Ono A,Matsuda A,Zhao J,Santi DV

doi

10.1093/nar/23.22.4677

subject

Has Abstract

pub_date

1995-11-25 00:00:00

pages

4677-82

issue

22

eissn

0305-1048

issn

1362-4962

pii

5a0441

journal_volume

23

pub_type

杂志文章
  • BCNTB bioinformatics: the next evolutionary step in the bioinformatics of breast cancer tissue banking.

    abstract::Here, we present an update of Breast Cancer Now Tissue Bank bioinformatics, a rich platform for the sharing, mining, integration and analysis of breast cancer data. Its modalities provide researchers with access to a centralised information gateway from which they can access a network of bioinformatic resources to que...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkx913

    authors: Gadaleta E,Pirrò S,Dayem Ullah AZ,Marzec J,Chelala C

    更新日期:2018-01-04 00:00:00

  • Major capsid reinforcement by a minor protein in herpesviruses and phage.

    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

    authors: Sae-Ueng U,Liu T,Catalano CE,Huffman JB,Homa FL,Evilevitch A

    更新日期:2014-08-01 00:00:00

  • DNA methylation in hereditary persistence of fetal hemoglobin (HPFH-2).

    abstract::Recent studies show that the region of DNA brought into close proximity to the fetal gamma-globin genes in deletional forms of hereditary persistence of fetal hemoglobin (HPFH) is selectively hypomethylated (and presumably active) in normal erythroid tissue. This region is normally located approximately 100 kilobases ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/15.13.5169

    authors: Poncz M,Sutton M,Delgrosso K,Schwartz E,Surrey S

    更新日期:1987-07-10 00:00:00

  • PatternQuery: web application for fast detection of biomacromolecular structural patterns in the entire Protein Data Bank.

    abstract::Well defined biomacromolecular patterns such as binding sites, catalytic sites, specific protein or nucleic acid sequences, etc. precisely modulate many important biological phenomena. We introduce PatternQuery, a web-based application designed for detection and fast extraction of such patterns. The application uses a...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkv561

    authors: Sehnal D,Pravda L,Svobodová Vařeková R,Ionescu CM,Koča J

    更新日期:2015-07-01 00:00:00

  • Glycyl-tRNA synthetase specifically binds to the poliovirus IRES to activate translation initiation.

    abstract::Adaptation to the host cell environment to efficiently take-over the host cell's machinery is crucial in particular for small RNA viruses like picornaviruses that come with only small RNA genomes and replicate exclusively in the cytosol. Their Internal Ribosome Entry Site (IRES) elements are specific RNA structures th...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gks182

    authors: Andreev DE,Hirnet J,Terenin IM,Dmitriev SE,Niepmann M,Shatsky IN

    更新日期:2012-07-01 00:00:00

  • Characterisation of a genomic clone covering the structural mouse MyoD1 gene and its promoter region.

    abstract::We have isolated the mouse MyoD1 gene flanked by its promoter region by screening a genomic library with synthetic oligonucleotides. The structural gene is interrupted by two G + C rich introns. Transfection of the cloned gene inserted into an expression vector converts fibroblasts to myoblasts. Sequence analysis of a...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/19.23.6433

    authors: Zingg JM,Alva GP,Jost JP

    更新日期:1991-12-11 00:00:00

  • Organization, structure and expression of murine interferon alpha genes.

    abstract::Using a human interferon-alpha probe we have isolated recombinant phages containing murine interferon-alpha (Mu IFN-alpha) genes from a genomic library. One of these phages contained two complete Mu IFN-alpha genes and part of a third gene. The insert of a second phage held two IFN genes. This indicates that the Mu IF...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/13.3.791

    authors: Zwarthoff EC,Mooren AT,Trapman J

    更新日期:1985-02-11 00:00:00

  • The essential transcription factor Reb1p interacts with the CLB2 UAS outside of the G2/M control region.

    abstract::Regulation of CLB2 is important both for completion of the normal vegetative cell cycle in Saccharomyces cerevisiae and for departure from the vegetative cell cycle upon nitrogen deprivation. Cell cycle-regulated transcription of CLB2 in the G2/M phase is known to be brought about by a set of proteins including Mcm1p,...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkg638

    authors: Van Slyke C,Grayhack EJ

    更新日期:2003-08-01 00:00:00

  • Transcriptional regulation of the apoAI gene by hepatic nuclear factor 4 in yeast.

    abstract::Hepatocyte Nuclear Factor 4 (HNF-4), a liver-enriched orphan receptor of the nuclear receptor superfamily, is required for the expression of a wide variety of liver-specific genes including apoAI. To explore the possibility that site A of the apoAI gene enhancer might also be the target for HNF-4 without the interfere...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/22.25.5665

    authors: Fuernkranz HA,Wang Y,Karathanasis SK,Mak P

    更新日期:1994-12-25 00:00:00

  • Development of a short-term, in vivo mutagenesis assay: the effects of methylation on the recovery of a lambda phage shuttle vector from transgenic mice.

    abstract::Transgenic mice suitable for the in vivo assay of suspected mutagens at the chromosome level have been constructed by stable integration of a lambda phage shuttle vector. The shuttle vector, which contains a beta-galactosidase (beta-gal) target gene, can be rescued from genomic DNA with in vitro packaging extracts. Mu...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/18.10.3007

    authors: Kohler SW,Provost GS,Kretz PL,Dycaico MJ,Sorge JA,Short JM

    更新日期:1990-05-25 00:00:00

  • The beta recombinase from the Streptococcal plasmid pSM 19035 represses its own transcription by holding the RNA polymerase at the promoter region.

    abstract::The beta protein encoded by the Streptococcus pyogenes plasmid pSM19035 is a site-specific recombinase involved in both resolution of plasmid multimers into monomers and DNA inversion. It has been proposed that the DNA region to which the beta recombinase binds to mediate recombination includes a promoter from which o...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/22.10.1855

    authors: Rojo F,Alonso JC

    更新日期:1994-05-25 00:00:00

  • Nucleotide sequence of an heterochromatic segment recognized by the antibodies to Z-DNA in fixed metaphase chromosomes.

    abstract::The purpose of this work was to analyse at the molecular level the DNA recognized by the antibodies to Z-DNA in in situ experiments. Antibodies to Z-DNA interact strongly with R-band positive heterochromatic segments of fixed metaphase chromosomes of Cebus (Viegas-Pequignot et al., 1983). These segments are constitute...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/14.8.3197

    authors: Malfoy B,Rousseau N,Vogt N,Viegas-Pequignot E,Dutrillaux B,Leng M

    更新日期:1986-04-25 00:00:00

  • Therapeutic target database update 2018: enriched resource for facilitating bench-to-clinic research of targeted therapeutics.

    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

    authors: Li YH,Yu CY,Li XX,Zhang P,Tang J,Yang Q,Fu T,Zhang X,Cui X,Tu G,Zhang Y,Li S,Yang F,Sun Q,Qin C,Zeng X,Chen Z,Chen YZ,Zhu F

    更新日期:2018-01-04 00:00:00

  • Sequence signal involved in the generation of an internally deleted defective interfering RNA from vesicular stomatitis virus.

    abstract::We have determined the nucleotide sequence at the 3' end of the vesicular stomatitis virus (VSV) polymerase or L gene and compared it to that obtained from a defective interfering particle (DI) RNA generated by this virus. The latter (DI 0.50) contains a large internal deletion within this gene. The deletion begins ex...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/10.21.6919

    authors: De BK,Perrault J

    更新日期:1982-11-11 00:00:00

  • Histone modifications influence mediator interactions with chromatin.

    abstract::The Mediator complex transmits activation signals from DNA bound transcription factors to the core transcription machinery. Genome wide localization studies have demonstrated that Mediator occupancy not only correlates with high levels of transcription, but that the complex also is present at transcriptionally silence...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkr551

    authors: Zhu X,Zhang Y,Bjornsdottir G,Liu Z,Quan A,Costanzo M,Dávila López M,Westholm JO,Ronne H,Boone C,Gustafsson CM,Myers LC

    更新日期:2011-10-01 00:00:00

  • Replication of a carcinogenic nitropyrene DNA lesion by human Y-family DNA polymerase.

    abstract::Nitrated polycyclic aromatic hydrocarbons are common environmental pollutants, of which many are mutagenic and carcinogenic. 1-Nitropyrene is the most abundant nitrated polycyclic aromatic hydrocarbon, which causes DNA damage and is carcinogenic in experimental animals. Error-prone translesion synthesis of 1-nitropyre...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gks1296

    authors: Kirouac KN,Basu AK,Ling H

    更新日期:2013-02-01 00:00:00

  • Adenine-guanine base pairing ribosomal RNA.

    abstract::Analyses of secondary structures proposed for ribosomal RNA's show that, of the different kinds of base pairs directly adjoining the ends of postulated double-helical regions, only A-G with A at the 5' end significantly exceeds the number expected for a random base distribution. An A(syn)-G(trans) hydrogen-bonded base...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/10.8.2701

    authors: Traub W,Sussman JL

    更新日期:1982-04-24 00:00:00

  • All three functional domains of the large ribosomal subunit protein L25 are required for both early and late pre-rRNA processing steps in Saccharomyces cerevisiae.

    abstract::Mutational analysis has shown that the integrity of the region in domain III of 25S rRNA that is involved in binding of ribosomal protein L25 is essential for the production of mature 25S rRNA in the yeast Saccharomyces cerevisiae. However, even structural alterations that do not noticeably affect recognition by L25, ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/29.24.5001

    authors: van Beekvelt CA,de Graaff-Vincent M,Faber AW,van't Riet J,Venema J,Raué HA

    更新日期:2001-12-15 00:00:00

  • 4EHP-independent repression of endogenous mRNAs by the RNA-binding protein GIGYF2.

    abstract::Initially identified as a factor involved in tyrosine kinase receptor signaling, Grb10-interacting GYF protein 2 (GIGYF2) has later been shown to interact with the 5' cap-binding protein 4EHP as part of a translation repression complex, and to mediate post-transcriptional repression of tethered reporter mRNAs. A curre...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gky198

    authors: Amaya Ramirez CC,Hubbe P,Mandel N,Béthune J

    更新日期:2018-06-20 00:00:00

  • The SV40 72 base repair repeat has a striking effect on gene expression both in SV40 and other chimeric recombinants.

    abstract::By introduction of recombinant plasmids into monkey CV1 cells, we have unambiguously demonstrated that sequences entirely within the 72 bp repeat, which is located upstream of the SV40 early region, are crucial for T-antigen expression in vivo. We have also shown that a DNA fragment containing the 72 bp repeat, insert...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/9.22.6047

    authors: Moreau P,Hen R,Wasylyk B,Everett R,Gaub MP,Chambon P

    更新日期:1981-11-25 00:00:00

  • H1a, an E. coli DNA-binding protein which accumulates in stationary phase, strongly compacts DNA in vitro.

    abstract::We characterize a component of the E. coli bacterial nucleoid H1a, which accumulates in stationary phase. This protein, identical with the major component of a plasmid-protein complex previously isolated in our laboratory, has a pI close to 7.5. Acrylamide gel electrophoresis and sedimentation in sucrose gradient have...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/12.13.5321

    authors: Spassky A,Rimsky S,Garreau H,Buc H

    更新日期:1984-07-11 00:00:00

  • Array-based analysis of genomic DNA methylation patterns of the tumour suppressor gene p16INK4A promoter in colon carcinoma cell lines.

    abstract::Aberrant DNA methylation at CpG dinucleotides can result in epigenetic silencing of tumour suppressor genes and represents one of the earliest events in tumourigenesis. To date, however, high-throughput tools that are capable of surveying the methylation status of multiple gene promoters have been restricted to a limi...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gni072

    authors: Mund C,Beier V,Bewerunge P,Dahms M,Lyko F,Hoheisel JD

    更新日期:2005-04-28 00:00:00

  • Single base extension (SBE) with proofreading polymerases and phosphorothioate primers: improved fidelity in single-substrate assays.

    abstract::Model single base extension (SBE) genotyping reactions with individual deoxy-, dideoxy- and acyclonucleoside triphosphates are monitored by MALDI-TOF mass spectrometry. Three non-proofreading DNA polymerases display remarkably high misincorporation (up to 64% of correct incorporation) when extending primers with singl...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gng007

    authors: Di Giusto D,King GC

    更新日期:2003-02-01 00:00:00

  • Genomic organization and transcription of the alpha beta heat shock DNA in Drosophila melanogaster.

    abstract::Previous studies have shown that (i) several RNAs induced by heat shock of Drosophila melanogaster cells are homologous to tandemly repeated alpha beta units found in cloned segments of D. melanogaster DNA, and (ii) the alpha beta sequences are present both at a major heat shock locus, 87Cl, and the chromocenter of po...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/9.20.5297

    authors: Lis JT,Ish-Horowicz D,Pinchin SM

    更新日期:1981-10-24 00:00:00

  • A novel activity of HMG domains: promotion of the triple-stranded complex formation between DNA containing (GGA/TCC)11 and d(GGA)11 oligonucleotides.

    abstract::The high mobility group protein (HMG)-box is a DNA-binding domain found in many proteins that bind preferentially to DNA of irregular structures in a sequence-independent manner and can bend the DNA. We show here that GST-fusion proteins of HMG domains from HMG1 and HMG2 promote a triple-stranded complex formation bet...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/24.23.4733

    authors: Suda T,Mishima Y,Takayanagi K,Asakura H,Odani S,Kominami R

    更新日期:1996-12-01 00:00:00

  • Complementary addressed modification of yeast tRNA Val 1 with alkylating derivative of d(pC-G)-A. The positions of the alkylated nucleotides and the course of the alkylation in the complex.

    abstract::Yeast tRNA Val 1 alkylation with 2', 3'-O-4-(N-2-chloroethyl-N-methylamino) benzylidene d(pC-G)-A proceeds at 20 degrees - 30 degrees C in the complementary complexes which are formed by d(pC-G)-A greater than RC1 binding to 3 sequences of tRNA Val 1 : psi-C-G58 in the T loop, C-G40 at the 3'-side of the anticodon loo...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/4.5.1609

    authors: Grineva NI,Karpova GG,Kuznetsova LM,Venkstern TV,Bayev AA

    更新日期:1977-01-01 00:00:00

  • The yeast POP2 gene encodes a nuclease involved in mRNA deadenylation.

    abstract::The major mRNA degradation pathway involves deadenylation of the target molecule followed by decapping and, finally, 5'-->3' exonuclease digestion of the mRNA body. While yeast factors involved in the decapping and exonuclease degradation steps have been identified, the nature of the factor(s) involved in the deadenyl...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/29.12.2448

    authors: Daugeron MC,Mauxion F,Séraphin B

    更新日期:2001-06-15 00:00:00

  • The optimal binding sequence of the Hox11 protein contains a predicted recognition core motif.

    abstract::HOX11 is a homeobox-containing oncogene of specific T-cell leukemias. We determined the DNA binding specificity of the Hox11 protein by using a novel technique of random oligonucleotide selection developed in this study. The optimal Hox11 binding sequence, GGCGGTAAGTGG, contained a core TAAGTG motif that is consistent...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/23.11.1928

    authors: Tang S,Breitman ML

    更新日期:1995-06-11 00:00:00

  • The localization of tRNA4Glu genes from Drosophila melanogaster by "in situ" hybridization.

    abstract::Transfer RNAGlu4 was isolated from Drosophila melanogaster by affinity chromatography. The tRNA was iodinated "in vitro" with Na[25I] and hybridized "in situ" to salivary gland chromosomes from Drosophila. Subsequent autoradiography allowed the localization of the genes for tRNAGlu4 to the right arm of the second chro...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/5.5.1465

    authors: Kubli E,Schmidt T

    更新日期:1978-05-01 00:00:00

  • ProPortal: a resource for integrated systems biology of Prochlorococcus and its phage.

    abstract::ProPortal (http://proportal.mit.edu/) is a database containing genomic, metagenomic, transcriptomic and field data for the marine cyanobacterium Prochlorococcus. Our goal is to provide a source of cross-referenced data across multiple scales of biological organization--from the genome to the ecosystem--embracing the f...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkr1022

    authors: Kelly L,Huang KH,Ding H,Chisholm SW

    更新日期:2012-01-01 00:00:00