A DNA cassette containing a trimerized SV40 polyadenylation signal which efficiently blocks spurious plasmid-initiated transcription.

Abstract:

:A head-to-tail trimer of the SV40 Bcl I-Bam H1 DNA fragment, specifying polyadenylation of RNA transcripts, was cloned as a cassette flanked by multiple restriction sites. Insertion of the trimer into several expression vectors efficiently prevented spurious expression of reporter genes resulting from transcriptional initiation in prokaryotic plasmid sequences in transfected mammalian cells.

journal_name

Biotechniques

journal_title

BioTechniques

authors

Maxwell IH,Harrison GS,Wood WM,Maxwell F

subject

Has Abstract

pub_date

1989-03-01 00:00:00

pages

276-80

issue

3

eissn

0736-6205

issn

1940-9818

journal_volume

7

pub_type

杂志文章
  • Cre stoplight: a red/green fluorescent reporter of Cre recombinase expression in living cells.

    abstract::The Cre/lox system is a powerful genetic tool with which to manipulate the genome. Here, we describe the development of a simple reporter system for Cre recombinase, called the Cre Stoplight. In the absence of Cre, the red fluorescent protein is expressed; when Cre catalyzes a recombination event, the green fluorescen...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/01315st03

    authors: Yang YS,Hughes TE

    更新日期:2001-11-01 00:00:00

  • Techniques in plant telomere biology.

    abstract::The role model systems have played in understanding telomere biology has been enormous, and understanding has rapidly transferred to human telomere research. Most work using model organisms to study telomerase and nontelomerase-based telomere-maintenance systems has centered on yeasts, ciliates, and insects. But it is...

    journal_title:BioTechniques

    pub_type: 杂志文章,评审

    doi:10.2144/05382RV01

    authors: Fajkus J,Sýkorová E,Leitch AR

    更新日期:2005-02-01 00:00:00

  • Tissue engineering tools for modulation of the immune response.

    abstract::Tissue engineering scaffolds have emerged as a powerful tool within regenerative medicine. These materials are being designed to create environments that promote regeneration through a combination of: (i) scaffold architecture, (ii) the use of scaffolds as vehicles for transplanting progenitor cells, and/or (iii) loca...

    journal_title:BioTechniques

    pub_type: 杂志文章,评审

    doi:10.2144/000113754

    authors: Boehler RM,Graham JG,Shea LD

    更新日期:2011-10-01 00:00:00

  • A novel vector for positive selection of inserts harboring an open reading frame by translational coupling.

    abstract::We have developed a novel vector pTCS, as a tool for efficient selection of open reading frame (ORF)-containing inserts. In pTCS clones containing an insert with an ORF a downstream marker gene (immE3, conferring resistance to colicin) is activated via translational coupling with the insert, and transformed cells can ...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/000112629

    authors: Ohashi-Kunihiro S,Yohda M,Masaki H,Machida M

    更新日期:2007-12-01 00:00:00

  • Application of cDNA arrays to monitor mRNA profiles in single preimplantation mouse embryos.

    abstract::Array technology is a widely used tool for gene expression profiling in various biological systems. However, the application of this method to mammalian preimplantation embryos is limited by the small amount of mRNA that can be extracted from a single embryo, which is not sufficient for array analysis. Here we report ...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/02332rr04

    authors: Brambrink T,Wabnitz P,Halter R,Klocke R,Carnwath J,Kues W,Wrenzycki C,Paul D,Niemann H

    更新日期:2002-08-01 00:00:00

  • Beta-lactamase: an ideal reporter system for monitoring gene expression in live eukaryotic cells.

    abstract::To gain insightful information about the mechanisms through which genes are activated and repressed requires gene reporter systems that are sensitive, robust, and cost-effective. Although numerous reporter gene technologies are commercially available, none are as sophisticated and user-friendly as beta-lactamase (BLA)...

    journal_title:BioTechniques

    pub_type: 杂志文章,评审

    doi:10.2144/000112292

    authors: Qureshi SA

    更新日期:2007-01-01 00:00:00

  • Fluorescent in situ hybridization employing the conventional NBT/BCIP chromogenic stain.

    abstract::In situ hybridization techniques typically employ chromogenic staining by enzymatic amplification to detect domains of gene expression. We demonstrate the previously unreported near infrared (NIR) fluorescence of the dark purple stain formed from the commonly used chromogens, nitro blue tetrazolium (NBT) and 5-bromo-4...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/000112476

    authors: Trinh le A,McCutchen MD,Bonner-Fraser M,Fraser SE,Bumm LA,McCauley DW

    更新日期:2007-06-01 00:00:00

  • Silencing of antiapoptotic survivin gene by multiple approaches of RNA interference technology.

    abstract::Silencing of mammalian gene expression by RNA interference (RNAi) technology can be achieved using small interfering RNA (siRNA) or short hairpin RNA (shRNA). However, the relative effectiveness of these two approaches is not known. It is also not clear whether gene-specific shRNA transcribed from an RNA polymerase II...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/04363RR01

    authors: Ling X,Li F

    更新日期:2004-03-01 00:00:00

  • Rapid regeneration of virus from cells infected with a retroviral vector.

    abstract::Recombinant retroviral vectors usually encode the genes of interest in place of the viral structural genes, which must be provided in trans. These viruses are therefore defective for replication: infected cells cannot produce progeny virus. However, in some cases it may be desirable to generate virus from an infected ...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Corbley MJ,Cherington V,Feig LA,Cooper GM,Roberts TM

    更新日期:1994-12-01 00:00:00

  • Patents and patent office resources in biotechnology.

    abstract::Patents play an increasingly important role in the dissemination of information in many fast moving fields such as biotechnology and semiconductors. Quite a few new developments are introduced as patents, and only later, if at all, do they find their way into the scientific literature. In spite of this, patents lack w...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Koniarek JP,Coleman KD

    更新日期:1988-02-01 00:00:00

  • Affinity as a tool in life science.

    abstract::The use of affinity-based tools has become invaluable as a platform for basic research and in the development of drugs and diagnostics. Applications include affinity chromatography and affinity tag fusions for efficient purification of proteins as well as methods to probe the protein network interactions on a whole-pr...

    journal_title:BioTechniques

    pub_type: 杂志文章,评审

    doi:10.2144/000112803

    authors: Uhlén M

    更新日期:2008-04-01 00:00:00

  • A rapid and simple method for labeling short DNA fragments using Taq polymerase.

    abstract::This report describes a new method for labeling PCR-generated short length (60-120 bp) double-stranded DNA fragments for use as hybridization probes. The method utilizes gene-specific primers identical to those for PCR generation of non-radioactive DNA fragments. Radioactive probes are synthesized by Taq DNA polymeras...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Mizobuchi M,Frohman LA

    更新日期:1992-03-01 00:00:00

  • TEV protease-mediated cleavage in Drosophila as a tool to analyze protein functions in living organisms.

    abstract::Drosophila provides a powerful experimental system to analyze gene functions in a multi-cellular organism. Here we describe an in vivo method that interferes with the integrity of selected proteins through site-specific cleavage in Drosophila. The technique is based on the highly specific seven-amino-acid recognition ...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/000112884

    authors: Harder B,Schomburg A,Pflanz R,Küstner K,Gerlach N,Schuh R

    更新日期:2008-05-01 00:00:00

  • A hemagglutinating substance in chitin.

    abstract::Chitin from crustacean shells has often been used to isolate and purify plant lectins that have an affinity for poly-N-acetylglucosamine (poly-GlcNAc). When we used washed chitin from crab shells as an affinity medium to isolate a lectin from Pinus strobus L. (eastern white pine) ovules, we found that a substance havi...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Whitmore FA

    更新日期:1992-02-01 00:00:00

  • Localization of trinucleotide repeat sequences in myotonic dystrophy cells using a single fluorochrome-labeled PNA probe.

    abstract::A labeled peptide nucleic acid (PNA) antisense probe was used to study the spatial distribution of triplet repeats (CTG) in human myotonic dystrophy (DM) cells by high-resolution fluorescence in situ hybridization (FISH). It was found that transcripts containing triplet repeats were present as a number of discrete foc...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/98243rr02

    authors: Taneja KL

    更新日期:1998-03-01 00:00:00

  • Self-seal reagent: evaporation control for molecular histology procedures without chambers, clips or fingernail polish.

    abstract::Sensitive nucleic acid based detection methods such as in situ PCR, in situ RT-PCR and PRINS have great potential in the areas of developmental biology, pathogenesis and diagnostics. However, control of evaporation from in situ reactions is critical to ensure reliable data. Self-Seal Reagent, a component added directl...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/97232pf01

    authors: Sullivan DE,Bobroski LE,Bagasra O,Finney M

    更新日期:1997-08-01 00:00:00

  • Preparation of amino acid mixtures for cell-free expression systems.

    abstract::Here we present a procedure for preparing amino acid mixtures--having both the desired composition and a physiological pH--at high concentrations for cell-free expression systems. Up to 2.1 mg/mL of active protein was synthesized in batch mode reactions with an all Escherichia coli cell-free expression system. Our met...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/000114249

    authors: Caschera F,Noireaux V

    更新日期:2015-01-01 00:00:00

  • Efficacy of an amphipathic oligopeptide to shuttle and release a cis-acting DNA decoy into human cells.

    abstract::We investigated the ability of an amphipathic oligopeptide to carry a synthetic dsDNA oligonucleotide inside human cells. The oligonucleotide was designed as a decoy binding site for the transcriptional activator of the methylguanine-DNA methyltransferase (MGMT) gene. The complex oligopeptide and decoy were administer...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/02321dd03

    authors: Citti L,Rovero P,Colombo MG,Mariani L,Poliseno L,Rainaldi G

    更新日期:2002-01-01 00:00:00

  • Automation of dideoxynucleotide DNA sequencing reactions using a robotic workstation.

    abstract::We report here a system for automation of the dideoxynucleotide DNA sequencing method. The system consists of a Beckman Biomek 1000 robotic workstation which has been modified by the addition of a thin heater/cooler block directly on the instrument table. The heater/cooler block, which is regulated by a user-specified...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Wilson RK,Yuen AS,Clark SM,Spence C,Arakelian P,Hood LE

    更新日期:1988-09-01 00:00:00

  • Specific suppression of false-positive signals in the product-enhanced reverse transcriptase assay.

    abstract::The recently developed ultrasensitive reverse transcriptase (RT) test involving a PCR step can detect minute amounts of RT in single retroviral particles and is 10(6) times more sensitive than conventional RT assays. We have found that different DNA-dependent DNA polymerases like DNA Polymerase I from Escherichia coli...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Lugert R,König H,Kurth R,Tönjes RR

    更新日期:1996-02-01 00:00:00

  • Multicolor luciferase assay system: one-step monitoring of multiple gene expressions with a single substrate.

    abstract::Reporter assays that use luciferase are widely employed for monitoring cellular events associated with gene expression. In general, firefly luciferase and Renilla luciferase are used for monitoring single gene expression. However, the expression of more than one gene cannot be monitored simultaneously by this system b...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/05386ST03

    authors: Nakajima Y,Kimura T,Sugata K,Enomoto T,Asakawa A,Kubota H,Ikeda M,Ohmiya Y

    更新日期:2005-06-01 00:00:00

  • Development of a PCR-based strategy for CYP2D6 genotyping including gene multiplication of worldwide potential use.

    abstract::There is growing consensus on the potential use of pharmacogenetics in clinical practice, and hopes have been expressed for application to the improvement of global health. However, two major challenges may lead to widening the "biotechnological gap" between the developing and the industrial world;first the unaffordab...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Dorado P,Cáceres MC,Pozo-Guisado E,Wong ML,Licinio J,Llerena A

    更新日期:2005-10-01 00:00:00

  • Sequential chemiluminescent detection of target DNAs without stripping and reprobing.

    abstract::We present a simple method for sequential chemiluminescent detections of two different DNA loci on a single Southern blot. First, an enzyme-linked DNA probe for a unique sequence is detected with a horse-radish peroxidase (HRP) substrate followed by the detection of another enzyme-linked DNA probe for a different uniq...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/99264st08

    authors: Reddy LV,DeSilva R,Handley RS,Schaap AP,Akhavan-Tafti H

    更新日期:1999-04-01 00:00:00

  • CDNA library construction from a small amount of RNA: adaptor-ligation approach for two-round cRNA amplification using T7 and SP6 RNA polymerases.

    abstract::In this study, we developed a method that allows cDNA library construction from a small amount of RNA without causing serious size bias in the resulting cDNA population. For this purpose, we adopted two-round cRNA amplification by T7 and SP6 RNA polymerases. The first-round cDNAs, flanked by the promoter sequences of ...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/05383RR01

    authors: Ohara R,Kikuno RF,Kitamura H,Ohara O

    更新日期:2005-03-01 00:00:00

  • Dispensing of very low volumes of ultra high viscosity alginate gels: a new tool for encapsulation of adherent cells and rapid prototyping of scaffolds and implants.

    abstract::We present a tool for dispensing very low volumes (20 nL or more) of ultra high viscosity (UHV) medical-grade alginate hydrogels. It uses a modified piezo-driven micrometering valve, integrated into a versatile system that allows fast prototyping of encapsulation procedures and scaffold production. Valves show excelle...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/000113014

    authors: Gepp MM,Ehrhart F,Shirley SG,Howitz S,Zimmermann H

    更新日期:2009-01-01 00:00:00

  • A method based on PCR for the construction of cDNA libraries and probes from small amounts of tissue.

    abstract::A PCR-based method is described for the production of cDNA libraries and total cDNA probes from a few milligrams of tissue. Using a model system, we show how a PCR library and PCR probes can be used to identify genes expressed at different levels in two tissues. Small amounts of tissue derived from two plants, one inf...

    journal_title:BioTechniques

    pub_type:

    doi:

    authors: Bertiol DJ,Smoker M,Brown AC,Jones MG,Burrows PR

    更新日期:1994-06-01 00:00:00

  • Optimizing leading edge F-actin labeling using multiple actin probes, fixation methods and imaging modalities.

    abstract::We systematically evaluated the performance and reliability of several widely used, commercially available actin-filament probes in a highly motile breast adenocarcinoma cell line to optimize the visualization of F-actin-rich dynamic lamellipodia. We evaluated four Phalloidin-fluorophores, two anti-actin antibodies, a...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/btn-2018-0112

    authors: DesMarais V,Eddy RJ,Sharma VP,Stone O,Condeelis JS

    更新日期:2019-03-01 00:00:00

  • Making cool drugs hot: isothermal titration calorimetry as a tool to study binding energetics.

    abstract::Characterization of the thermodynamics of binding interactions is important in improving our understanding of bimolecular recognition and forms an essential part of the rational drug design process. Isothermal titration calorimetry (ITC) is rapidly becoming established as the method of choice for undertaking such stud...

    journal_title:BioTechniques

    pub_type: 杂志文章,评审

    doi:

    authors: Holdgate GA

    更新日期:2001-07-01 00:00:00

  • Elimination of false positives that arise in using the two-hybrid system.

    abstract::The two-hybrid system is a genetic method to identify proteins that interact with a specific target protein, which is expressed in yeast as a hybrid with a DNA-binding domain. Use of this method entails transforming yeast, both with this DNA-binding domain hybrid and with a library of activation domain hybrids, follow...

    journal_title:BioTechniques

    pub_type:

    doi:

    authors: Bartel P,Chien CT,Sternglanz R,Fields S

    更新日期:1993-06-01 00:00:00

  • Assessment of three-dimensional biofilm structure using an optical microscope.

    abstract::A method allowing the evaluation of the structure and the calculation of the volume of a biofilm, using an optical microscope, is proposed based on the linear relation between the intensity of a pixel in biofilm images grabbed on the x-y plane and the corresponding number of cells in the z direction, which allows the ...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/000112403

    authors: de Carvalho CC,da Fonseca MM

    更新日期:2007-05-01 00:00:00