A DNA fragment with an alpha-phosphorothioate nucleotide at one end is asymmetrically blocked from digestion by exonuclease III and can be replicated in vivo.

Abstract:

:2'-Deoxyadenosine 5'-O-(1-thiotriphosphate) (dATP[alpha S]) was introduced into the 3' ends of DNA restriction fragments with Escherichia coli DNA polymerase I to give phosphorothioate internucleotide linkages. Such "capped" 3' ends were found to be resistant to exonuclease III digestion. Moreover, the resistance to digestion is great enough that, under conditions used by us, just one strand of a double helix is digested by exonuclease III when a cap is placed at only one end; when digestion is carried to completion, this results in production of intact single strands. When digestion with exonuclease III is limited and is followed by S1 nuclease treatment, double-stranded DNA fragments asymmetrically shortened from just one side are produced. In this was thousands of nucleotides can be selectively removed from one end of a restriction fragment. In vitro introduction of phosphorothioate linkages into one end of a linearized replicative plasmid, followed by exonuclease III and S1 nuclease treatments, gives rise to truncated forms that, upon circularization by blunt-end ligation, transform E. coli and replicate in vivo.

authors

Putney SD,Benkovic SJ,Schimmel PR

doi

10.1073/pnas.78.12.7350

subject

Has Abstract

pub_date

1981-12-01 00:00:00

pages

7350-4

issue

12

eissn

0027-8424

issn

1091-6490

journal_volume

78

pub_type

杂志文章
  • Imaging Tetrahymena ribozyme splicing activity in single live mammalian cells.

    abstract::Tetrahymena ribozymes hold promise for repairing genetic disorders but are largely limited by their modest splicing efficiency and low production of final therapeutic proteins. Ribozyme evolution in intact living mammalian cells would greatly facilitate the discovery of new ribozyme variants with high in vivo activity...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.2036553100

    authors: Hasegawa S,Jackson WC,Tsien RY,Rao J

    更新日期:2003-12-09 00:00:00

  • Rate enhancement of reconstitution of glyceraldehyde-3-phosphate dehydrogenase by a covalently bound coenzyme analog.

    abstract::Kinetic analysis of the in vitro reconstitution of glyceraldehyde-3-phosphate dehydrogenase [D-glyceraldehyde-3-phosphate:NAD+ oxidoreductase (phosphorylating), EC 1.2.1.12] from yeast showed that both oxidized and reduced coenzyme enhance the transconformation reaction, which is rate limiting in the sequential foldin...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.77.4.1966

    authors: Jaenicke R,Krebs H,Rudolph R,Woenckhaus C

    更新日期:1980-04-01 00:00:00

  • In vivo and in vitro observations on laminin production by Schwann cells.

    abstract::Recent reports on the dystrophic mouse mutant suggest that the prominent extracellular matrix component of peripheral nerve tissues plays an important role in peripheral nerve development. We have examined the disposition of two prominent extracellular matrix components, fibronectin and laminin, both in mature periphe...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.80.12.3850

    authors: Cornbrooks CJ,Carey DJ,McDonald JA,Timpl R,Bunge RP

    更新日期:1983-06-01 00:00:00

  • ATP-dependent structural change of the eukaryotic clamp-loader protein, replication factor C.

    abstract::The eukaryotic DNA sliding clamp that keeps DNA polymerase engaged at a replication fork, called proliferating cell nuclear antigen (PCNA), is loaded onto the 3' ends of primer DNA through its interaction with a heteropentameric protein complex called replication factor C (RFC). The ATPase activity of RFC is necessary...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.97.26.14127

    authors: Shiomi Y,Usukura J,Masamura Y,Takeyasu K,Nakayama Y,Obuse C,Yoshikawa H,Tsurimoto T

    更新日期:2000-12-19 00:00:00

  • The specific direct interaction of helper T cells and antigen-presenting B cells.

    abstract::Cell couples have been formed by mixing an antigen- and Ia-specific cloned helper T-cell line with a B-cell hybridoma presenting the antigen. By immunofluorescence observations, we have shown that the microtubule-organizing center (MTOC) inside the helper T cell, but not in the bound antigen-presenting cell, becomes o...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.83.16.6080

    authors: Kupfer A,Swain SL,Janeway CA Jr,Singer SJ

    更新日期:1986-08-01 00:00:00

  • DNA strand breakage, activation of poly (ADP-ribose) synthetase, and cellular energy depletion are involved in the cytotoxicity of macrophages and smooth muscle cells exposed to peroxynitrite.

    abstract::The free radicals nitric oxide and superoxide anion react to form peroxynitrite (ONOO-), a highly toxic oxidant species. In vivo formation of ONOO- has been demonstrated in shock and inflammation. Herein we provide evidence that cytotoxicity in cells exposed to ONOO- is mediated by DNA strand breakage and the subseque...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.93.5.1753

    authors: Szabó C,Zingarelli B,O'Connor M,Salzman AL

    更新日期:1996-03-05 00:00:00

  • A general model of invariant chain association with class II major histocompatibility complex proteins.

    abstract::The binding of invariant chain to major histocompatibility complex (MHC) proteins is an important step in processing of MHC class II proteins and in antigen presentation. The question of how invariant chain can bind to all MHC class II proteins is central to understanding these processes. We have employed molecular mo...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.92.18.8269

    authors: Lee C,McConnell HM

    更新日期:1995-08-29 00:00:00

  • Catalysis of amide synthesis by RNA phosphodiester and hydroxyl groups.

    abstract::The functional groups found among the RNA bases and in the phosphoribose backbone represent a limited repertoire from which to construct a ribozyme active site. This work investigates the possibility that simple RNA phosphodiester and hydroxyl functional groups could catalyze amide bond synthesis. Reaction of amine gr...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.212527799

    authors: Chamberlin SI,Merino EJ,Weeks KM

    更新日期:2002-11-12 00:00:00

  • Germ-line expression of an oncogenic erbB2 allele confers resistance to erbB2-induced mammary tumorigenesis.

    abstract::We have previously shown that mammary epithelial specific expression of the activated erbB2 allele under the control of the endogenous promoter in mice resulted in the formation of mammary adenocarcinomas. To assess whether mammary tumorigenesis in this model is influenced by the developmental window of expression, we...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0306802101

    authors: Andrechek ER,Hardy WR,Laing MA,Muller WJ

    更新日期:2004-04-06 00:00:00

  • Relationships between the responses of triglyceride-rich lipoproteins in blood plasma containing apolipoproteins B-48 and B-100 to a fat-containing meal in normolipidemic humans.

    abstract::The concentration of triglyceride-rich lipoproteins containing apolipoprotein (apo) B-48 (chylomicrons) and apo B-100 (very low density lipoproteins) was measured in blood plasma of healthy young men after an ordinary meal containing one-third of daily energy and fat. Plasma obtained in the postabsorptive state and at...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.90.5.2069

    authors: Schneeman BO,Kotite L,Todd KM,Havel RJ

    更新日期:1993-03-01 00:00:00

  • Some structural and antigenic properties of intracisternal A particles occurring in mouse tumors (complement fixation-immunodiffusion-neuroblastoma-plasma-cell tumor).

    abstract::Intracisternal A-particles were isolated from three different myeloma lines in BALB/c mice and from cultured neuroblastoma cells of A/J origin. All preparations contained a major structural protein with an apparent molecular weight near 70,000 as estimated by electrophoretic mobility in sodium dodecyl sulfate-containi...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.69.1.218

    authors: Kuff EL,Leuders KK,Ozer HL,Wivel NA

    更新日期:1972-01-01 00:00:00

  • Fibulin-4 exerts a dual role in LTBP-4L-mediated matrix assembly and function.

    abstract::Elastogenesis is a hierarchical process by which cells form functional elastic fibers, providing elasticity and the ability to regulate growth factor bioavailability in tissues, including blood vessels, lung, and skin. This process requires accessory proteins, including fibulin-4 and -5, and latent TGF binding protein...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1901048116

    authors: Kumra H,Nelea V,Hakami H,Pagliuzza A,Djokic J,Xu J,Yanagisawa H,Reinhardt DP

    更新日期:2019-10-08 00:00:00

  • Rapid typing of tandemly repeated hypervariable loci by the polymerase chain reaction: application to the apolipoprotein B 3' hypervariable region.

    abstract::The 3' flanking region of the apolipoprotein B (apoB) gene contains a hypervariable region consisting of a variable number of tandemly repeated short A + T-rich DNA sequences (VNTRs). We present a general method that utilizes the polymerase chain reaction to rapidly and accurately type this and other VNTR loci. We use...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.86.1.212

    authors: Boerwinkle E,Xiong WJ,Fourest E,Chan L

    更新日期:1989-01-01 00:00:00

  • Extent of N-terminal methionine excision from Escherichia coli proteins is governed by the side-chain length of the penultimate amino acid.

    abstract::In a significant fraction of the Escherichia coli cytosolic proteins, the N-terminal methionine residue incorporated during the translation initiation step is excised. The N-terminal methionine excision is catalyzed by methionyl-aminopeptidase (MAP). Previous studies have suggested that the action of this enzyme could...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.86.21.8247

    authors: Hirel PH,Schmitter MJ,Dessen P,Fayat G,Blanquet S

    更新日期:1989-11-01 00:00:00

  • Widespread establishment and regulatory impact of Alu exons in human genes.

    abstract::The Alu element has been a major source of new exons during primate evolution. Thousands of human genes contain spliced exons derived from Alu elements. However, identifying Alu exons that have acquired genuine biological functions remains a major challenge. We investigated the creation and establishment of Alu exons ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1012834108

    authors: Shen S,Lin L,Cai JJ,Jiang P,Kenkel EJ,Stroik MR,Sato S,Davidson BL,Xing Y

    更新日期:2011-02-15 00:00:00

  • Inheritance of Trans Chromosomal Methylation patterns from Arabidopsis F1 hybrids.

    abstract::Hybridization in plants leads to transinteractions between the parental genomes and epigenomes that can result in changes to both 24 nt siRNA and cytosine methylation ((m)C) levels in the hybrid. In Arabidopsis the principle processes altering the hybrid methylome are Trans Chromosomal Methylation (TCM) and Trans Chro...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1323656111

    authors: Greaves IK,Groszmann M,Wang A,Peacock WJ,Dennis ES

    更新日期:2014-02-04 00:00:00

  • Proteins of Escherichia coli come in sizes that are multiples of 14 kDa: domain concepts and evolutionary implications.

    abstract::Initial attempts to correlate the distribution of gene density (number of gene loci per unit length on the linkage map) with the distribution of lengths of coding sequences have led to the observation that 46% of approximately 1000 sampled proteins in Escherichia coli have molecular masses of n X 14,000 +/- 2500 dalto...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.83.5.1198

    authors: Savageau MA

    更新日期:1986-03-01 00:00:00

  • Eddy current and coupled landscapes for nonadiabatic and nonequilibrium complex system dynamics.

    abstract::Physical and biological systems are often involved with coupled processes of different time scales. In the system with electronic and atomic motions, for example, the interplay between the atomic motion along the same energy landscape and the electronic hopping between different landscapes is critical: the system beha...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1305604110

    authors: Zhang K,Sasai M,Wang J

    更新日期:2013-09-10 00:00:00

  • Redirected infection of directly biotinylated recombinant adenovirus vectors through cell surface receptors and antigens.

    abstract::The inability of adenovirus to infect primitive hematopoietic cells presents an obstacle to the use of adenovirus vectors for gene transfer to these cell types. Therefore, expanding the tropism of adenovirus vectors to unique cell surface antigens would be an important development for gene therapy protocols. In this s...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.96.16.8855

    authors: Smith JS,Keller JR,Lohrey NC,McCauslin CS,Ortiz M,Cowan K,Spence SE

    更新日期:1999-08-03 00:00:00

  • Talin is phosphorylated on tyrosine in chicken embryo fibroblasts transformed by Rous sarcoma virus.

    abstract::We have examined the extent of tyrosine phosphorylation of talin, a component of the cytoskeleton localized in the focal adhesions and, therefore, a potential substrate of p60v-src, the transforming protein of Rous sarcoma virus. p60v-src is a tyrosine kinase that induces high levels of phosphotyrosine and the disorga...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.83.15.5507

    authors: Pasquale EB,Maher PA,Singer SJ

    更新日期:1986-08-01 00:00:00

  • Therapeutic cancer vaccines: using unique antigens.

    abstract::A decade ago, it seemed rational that our rapidly increasing knowledge of the molecular identities of tumor antigens and a deeper understanding of basic immunology would point the way to an effective therapeutic cancer vaccine. Significant progress has been made, but we do not yet have a cancer vaccine that can reliab...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章,评审

    doi:10.1073/pnas.0404839101

    authors: Lewis JJ

    更新日期:2004-10-05 00:00:00

  • Abrogation of T cell quiescence characterizes patients at high risk for multiple sclerosis after the initial neurological event.

    abstract::Clinically isolated syndrome (CIS) refers to the earliest clinical manifestation of multiple sclerosis (MS). Currently there are no prognostic biological markers that accurately predict conversion of CIS to clinically definite MS (CDMS). Furthermore, the earliest molecular events in MS are still unknown. We used micro...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0805065105

    authors: Corvol JC,Pelletier D,Henry RG,Caillier SJ,Wang J,Pappas D,Casazza S,Okuda DT,Hauser SL,Oksenberg JR,Baranzini SE

    更新日期:2008-08-19 00:00:00

  • Conformational transitions in DNA polymerase I revealed by single-molecule FRET.

    abstract::The remarkable fidelity of most DNA polymerases depends on a series of early steps in the reaction pathway which allow the selection of the correct nucleotide substrate, while excluding all incorrect ones, before the enzyme is committed to the chemical step of nucleotide incorporation. The conformational transitions t...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0910909107

    authors: Santoso Y,Joyce CM,Potapova O,Le Reste L,Hohlbein J,Torella JP,Grindley ND,Kapanidis AN

    更新日期:2010-01-12 00:00:00

  • The base pair-scale diffusion of nucleosomes modulates binding of transcription factors.

    abstract::The structure of promoter chromatin determines the ability of transcription factors (TFs) to bind to DNA and therefore has a profound effect on the expression levels of genes. However, the role of spontaneous nucleosome movements in this process is not fully understood. Here, we developed a single-molecule optical twe...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1815424116

    authors: Rudnizky S,Khamis H,Malik O,Melamed P,Kaplan A

    更新日期:2019-06-18 00:00:00

  • Activation of the hippocampus in normal humans: a functional anatomical study of memory.

    abstract::We studied regional cerebral blood flow using the H2(15)O method while normal subjects performed four similar tasks involving three-letter word beginnings (stems). Prior to each task, subjects studied a list of words. Local blood flow was then monitored during a 40-sec period while subjects (i) silently viewed word st...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.89.5.1837

    authors: Squire LR,Ojemann JG,Miezin FM,Petersen SE,Videen TO,Raichle ME

    更新日期:1992-03-01 00:00:00

  • Specific aneusomies in Chinese hamster cells at different stages of neoplastic transformation, initiated by nitrosomethylurea.

    abstract::Aneuploidy is ubiquitous in cancer, and its phenotypes are inevitably dominant and abnormal. In view of these facts we recently proposed that aneuploidy is sufficient for carcinogenesis generating cancer-specific aneusomies via a chain reaction of autocatalytic aneuploidizations. According to this hypothesis a carcino...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.251670699

    authors: Fabarius A,Willer A,Yerganian G,Hehlmann R,Duesberg P

    更新日期:2002-05-14 00:00:00

  • Molecular cloning of the pheromone biosynthesis-activating neuropeptide in Helicoverpa zea.

    abstract::Pheromone biosynthesis-activating neuropeptide (PBAN) regulates sex pheromone biosynthesis in female Helicoverpa (Heliothis) zea. Two oligonucleotide probes representing two overlapping amino acid regions of PBAN were used to screen 2.5 x 10(5) recombinant plaques, and a positive recombinant clone was isolated. Sequen...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.89.1.142

    authors: Davis MT,Vakharia VN,Henry J,Kempe TG,Raina AK

    更新日期:1992-01-01 00:00:00

  • Solar modulation of Little Ice Age climate in the tropical Andes.

    abstract::The underlying causes of late-Holocene climate variability in the tropics are incompletely understood. Here we report a 1,500-year reconstruction of climate history and glaciation in the Venezuelan Andes using lake sediments. Four glacial advances occurred between anno Domini (A.D.) 1250 and 1810, coincident with sola...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0603118103

    authors: Polissar PJ,Abbott MB,Wolfe AP,Bezada M,Rull V,Bradley RS

    更新日期:2006-06-13 00:00:00

  • Culling experiments demonstrate size-class specific biomass increases with mortality.

    abstract::Size-selective mortality inevitably leads to a decrease in population density and exerts a direct negative effect on targeted size classes. But density and population size structure are also shaped by food-dependent processes, such as individual growth, maturation, and reproduction. Mortality relaxes competition and t...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0808279106

    authors: Schröder A,Persson L,de Roos AM

    更新日期:2009-02-24 00:00:00

  • Loss of NFAT5 results in renal atrophy and lack of tonicity-responsive gene expression.

    abstract::The transcription factor NFAT5/TonEBP, a member of the NFAT/Rel family of transcription factors, has been implicated in diverse cellular responses, including the response to osmotic stress, integrin-dependent cell migration, T cell activation, and the Ras pathway in Drosophila. To clarify the in vivo role of NFAT5, we...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0308703100

    authors: López-Rodríguez C,Antos CL,Shelton JM,Richardson JA,Lin F,Novobrantseva TI,Bronson RT,Igarashi P,Rao A,Olson EN

    更新日期:2004-02-24 00:00:00