From Gateway to MultiSite Gateway in one recombination event.


BACKGROUND:Invitrogen Gateway technology exploits the integrase/att site-specific recombination system for directional cloning of PCR products and the subsequent subcloning into destination vectors. One or three DNA segments can be cloned using Gateway or MultiSite Gateway respectively. A vast number of single-site Gateway destination vectors have been created while MultiSite Gateway is limited to few destination vectors and therefore to few applications. The aim of this work was to make the MultiSite Gateway technology available for multiple biological purposes. RESULTS:We created a construct, pDONR-R4-R3, to easily convert any available Gateway destination vector to a MultiSite Gateway vector in a single recombination reaction. In addition, we designed pDONR-R4-R3 so that DNA fragments already cloned upstream or downstream of the Gateway cassette in the original destination vectors can still be utilized for promoter-gene or translational fusions after the conversion. CONCLUSION:Our tool makes MultiSite Gateway a more widely accessible technology and expands its applications by exploiting all the features of the Gateway vectors already available.


BMC Mol Biol


BMC molecular biology


Magnani E,Bartling L,Hake S




Has Abstract


2006-12-06 00:00:00










  • RNF168, a new RING finger, MIU-containing protein that modifies chromatin by ubiquitination of histones H2A and H2AX.

    abstract:BACKGROUND:Modulation of chromatin structure has emerged as a critical molecular device to control gene expression. Histones undergo different post-translational modifications that increase chromatin accessibility to a number of regulatory factors. Among them, histone ubiquitination appears relevant in nuclear processe...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Pinato S,Scandiuzzi C,Arnaudo N,Citterio E,Gaudino G,Penengo L

    更新日期:2009-06-05 00:00:00

  • The yfhQ gene of Escherichia coli encodes a tRNA:Cm32/Um32 methyltransferase.

    abstract:BACKGROUND:Naturally occurring tRNAs contain numerous modified nucleosides. They are formed by enzymatic modification of the primary transcripts during the complex RNA maturation process. In model organisms Escherichia coli and Saccharomyces cerevisiae most enzymes involved in this process have been identified. Interes...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Purta E,van Vliet F,Tkaczuk KL,Dunin-Horkawicz S,Mori H,Droogmans L,Bujnicki JM

    更新日期:2006-07-18 00:00:00

  • Deletion of the cruciform binding domain in CBP/14-3-3 displays reduced origin binding and initiation of DNA replication in budding yeast.

    abstract:BACKGROUND:Initiation of eukaryotic DNA replication involves many protein-protein and protein-DNA interactions. We have previously shown that 14-3-3 proteins bind cruciform DNA and associate with mammalian and yeast replication origins in a cell cycle dependent manner. RESULTS:By expressing the human 14-3-3epsilon, as...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Yahyaoui W,Callejo M,Price GB,Zannis-Hadjopoulos M

    更新日期:2007-04-12 00:00:00

  • Preliminary molecular characterization of the human pathogen Angiostrongylus cantonensis.

    abstract:BACKGROUND:Human angiostrongyliasis is an emerging food-borne public health problem, with the number of cases increasing worldwide, especially in mainland China. Angiostrongylus cantonensis is the causative agent of this severe disease. However, little is known about the genetics and basic biology of A. cantonensis. R...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: He H,Cheng M,Yang X,Meng J,He A,Zheng X,Li Z,Guo P,Pan Z,Zhan X

    更新日期:2009-10-25 00:00:00

  • Single and combinatorial chromatin coupling events underlies the function of transcript factor Krüppel-like factor 11 in the regulation of gene networks.

    abstract:BACKGROUND:Krüppel-like factors (KLFs) are a group of master regulators of gene expression conserved from flies to human. However, scant information is available on either the mechanisms or functional impact of the coupling of KLF proteins to chromatin remodeling machines, a deterministic step in transcriptional regula...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Calvo E,Grzenda A,Lomberk G,Mathison A,Iovanna J,Urrutia R

    更新日期:2014-05-25 00:00:00

  • RAD51 paralogs promote homology-directed repair at diversifying immunoglobulin V regions.

    abstract:BACKGROUND:Gene conversion depends upon the same factors that carry out more general process of homologous recombination, including homologous gene targeting and recombinational repair. Among these are the RAD51 paralogs, conserved factors related to the key recombination factor, RAD51. In chicken and other fowl, gene ...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Ordinario EC,Yabuki M,Handa P,Cummings WJ,Maizels N

    更新日期:2009-10-28 00:00:00

  • Post-translational generation of constitutively active cores from larger phosphatases in the malaria parasite, Plasmodium falciparum: implications for proteomics.

    abstract:BACKGROUND:Although the complete genome sequences of a large number of organisms have been determined, the exact proteomes need to be characterized. More specifically, the extent to which post-translational processes such as proteolysis affect the synthesized proteins has remained unappreciated. We examined this issue ...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Kumar R,Musiyenko A,Oldenburg A,Adams B,Barik S

    更新日期:2004-07-01 00:00:00

  • Accumulation of large non-circular forms of the chromosome in recombination-defective mutants of Escherichia coli.

    abstract:BACKGROUND:Double-strand breakage of chromosomal DNA is obviously a serious threat to cells because various activities of the chromosome depend on its integrity. However, recent experiments suggest that such breakage may occur frequently during "normal" growth in various organisms - from bacteria through vertebrates, p...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Handa N,Kobayashi I

    更新日期:2003-04-28 00:00:00

  • Regulation of poly(ADP-ribose) polymerase-1 (PARP-1) gene expression through the post-translational modification of Sp1: a nuclear target protein of PARP-1.

    abstract:BACKGROUND:Poly(ADP-ribose) polymerase-1 (PARP-1) is a nuclear enzyme that plays critical functions in many biological processes, including DNA repair and gene transcription. The main function of PARP-1 is to catalyze the transfer of ADP-ribose units from nicotinamide adenine dinucleotide (NAD+) to a large array of acc...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Zaniolo K,Desnoyers S,Leclerc S,Guérin SL

    更新日期:2007-10-25 00:00:00

  • L1CAM expression in endometrial carcinomas is regulated by usage of two different promoter regions.

    abstract:BACKGROUND:The L1 cell adhesion molecule (L1CAM) was originally identified as a neural adhesion molecule involved in axon guidance. In many human epithelial carcinomas L1CAM is overexpressed and thereby augments cell motility, invasion and metastasis formation. L1CAM positive carcinomas are associated with bad prognosi...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Pfeifer M,Schirmer U,Geismann C,Schäfer H,Sebens S,Altevogt P

    更新日期:2010-08-27 00:00:00

  • Genetic correction of splice site mutation in purified and enriched myoblasts isolated from mdx5cv mice.

    abstract:BACKGROUND:Duchenne Muscular Dystrophy (DMD) is an X-linked genetic disorder that results in the production of a dysfunctional form of the protein, dystrophin. The mdx5cv mouse is a model of DMD in which a point mutation in exon 10 of the dystrophin gene creates an artificial splice site. As a result, a 53 base pair de...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Maguire K,Suzuki T,DiMatteo D,Parekh-Olmedo H,Kmiec E

    更新日期:2009-02-23 00:00:00

  • Sequence periodicity of Escherichia coli is concentrated in intergenic regions.

    abstract:BACKGROUND:Sequence periodicity with a period close to the DNA helical repeat is a very basic genomic property. This genomic feature was demonstrated for many prokaryotic genomes. The Escherichia coli sequences display the period close to 11 base pairs. RESULTS:Here we demonstrate that practically only ApA/TpT dinucle...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Hosid S,Trifonov EN,Bolshoy A

    更新日期:2004-08-26 00:00:00

  • Critical evaluation of methods used to determine amplification efficiency refutes the exponential character of real-time PCR.

    abstract:BACKGROUND:The challenge of determining amplification efficiency has long been a predominant aspect of implementing real-time qPCR, playing a critical role in the accuracy and reliability that can be achieved. Based upon analysis of amplification profile position, standard curves are currently the gold standard for amp...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Rutledge RG,Stewart D

    更新日期:2008-10-30 00:00:00

  • Validation of reference genes for quantitative expression analysis by real-time RT-PCR in Saccharomyces cerevisiae.

    abstract:BACKGROUND:Real-time RT-PCR is the recommended method for quantitative gene expression analysis. A compulsory step is the selection of good reference genes for normalization. A few genes often referred to as HouseKeeping Genes (HSK), such as ACT1, RDN18 or PDA1 are among the most commonly used, as their expression is a...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Teste MA,Duquenne M,François JM,Parrou JL

    更新日期:2009-10-30 00:00:00

  • The major transcription initiation site of the p27Kip1 gene is conserved in human and mouse and produces a long 5'-UTR.

    abstract:BACKGROUND:The cyclin-dependent kinase inhibitor p27Kip1 is essential for proper control of cell cycle progression. The levels of p27Kip1 are regulated by several mechanisms including transcriptional and translational controls. In order to delineate the molecular details of these regulatory mechanisms it is important t...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Coleman J,Hawkinson M,Miskimins R,Miskimins WK

    更新日期:2001-01-01 00:00:00

  • Reference loci for RT-qPCR analysis of differentiating human embryonic stem cells.

    abstract:BACKGROUND:Selecting stably expressed reference genes is essential for proper reverse transcription quantitative polymerase chain reaction gene expression analysis. However, this choice is not always straightforward. In the case of differentiating human embryonic stem (hES) cells, differentiation itself introduces chan...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Vossaert L,O'Leary T,Van Neste C,Heindryckx B,Vandesompele J,De Sutter P,Deforce D

    更新日期:2013-09-12 00:00:00

  • Translation initiation region sequence preferences in Escherichia coli.

    abstract:BACKGROUND:The mRNA translation initiation region (TIR) comprises the initiator codon, Shine-Dalgarno (SD) sequence and translational enhancers. Probably the most abundant class of enhancers contains A/U-rich sequences. We have tested the influence of SD sequence length and the presence of enhancers on the efficiency o...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Vimberg V,Tats A,Remm M,Tenson T

    更新日期:2007-10-31 00:00:00

  • Genome-wide identification of brain miRNAs in response to high-intensity intermittent swimming training in Rattus norvegicus by deep sequencing.

    abstract:BACKGROUND:Physical exercise can improve brain function by altering brain gene expression. The expression mechanisms underlying the brain's response to exercise still remain unknown. miRNAs as vital regulators of gene expression may be involved in regulation of brain genes in response to exercise. However, as yet, very...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Zhao Y,Zhang A,Wang Y,Hu S,Zhang R,Qian S

    更新日期:2019-01-15 00:00:00

  • Comparative in silico analysis identifies bona fide MyoD binding sites within the Myocyte stress 1 gene promoter.

    abstract:BACKGROUND:Myocyte stress 1 (MS1) is a striated muscle actin binding protein required for the muscle specific activity of the evolutionary ancient myocardin related transcription factor (MRTF)/serum response factor (SRF) transcriptional pathway. To date, little is known about the molecular mechanisms that govern skelet...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Ounzain S,Dacwag CS,Samani NJ,Imbalzano AN,Chong NW

    更新日期:2008-05-19 00:00:00

  • A modification of Representational Difference Analysis, with application to the cloning of a candidate in the Reelin signalling pathway.

    abstract:BACKGROUND:cDNA-RDA is one of the subtractive cloning techniques used to isolate differentially expressed genes between two complex cDNA populations. In the present study we present a modification of the protocol described by Hubank and Schatz. RESULTS:In the post-hybridization mix, the 5'-ends of homoduplexes of inte...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Kuvbachieva AA,Goffinet AM

    更新日期:2002-04-24 00:00:00

  • What is normal? Next generation sequencing-driven analysis of the human circulating miRNAOme.

    abstract:BACKGROUND:MicroRNAs (miRNAs) are short non-protein-coding RNA species that have a regulatory function in modulating protein translation and degradation of specific mRNAs. MicroRNAs are estimated to target approximately 60% of all human mRNAs and are associated with the regulation of all physiological processes. Simila...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Tonge DP,Gant TW

    更新日期:2016-02-09 00:00:00

  • Housekeeping while brain's storming Validation of normalizing factors for gene expression studies in a murine model of traumatic brain injury.

    abstract:BACKGROUND:Traumatic brain injury models are widely studied, especially through gene expression, either to further understand implied biological mechanisms or to assess the efficiency of potential therapies. A large number of biological pathways are affected in brain trauma models, whose elucidation might greatly benef...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Rhinn H,Marchand-Leroux C,Croci N,Plotkine M,Scherman D,Escriou V

    更新日期:2008-07-08 00:00:00

  • Ataxia telangiectasia mutated (ATM) interacts with p400 ATPase for an efficient DNA damage response.

    abstract:BACKGROUND:Ataxia telangiectasia mutated (ATM) and TRRAP proteins belong to the phosphatidylinositol 3-kinase-related kinase family and are involved in DNA damage repair and chromatin remodeling. ATM is a checkpoint kinase that is recruited to sites of DNA double-strand breaks where it phosphorylates a diverse range of...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Smith RJ,Savoian MS,Weber LE,Park JH

    更新日期:2016-11-04 00:00:00

  • Analysis of the regulation of fatty acid binding protein 7 expression in human renal carcinoma cell lines.

    abstract:BACKGROUND:Improving the treatment of renal cell carcinoma (RCC) will depend on the development of better biomarkers for predicting disease progression and aiding the design of appropriate therapies. One such marker may be fatty acid binding protein 7 (FABP7), also known as B-FABP and BLBP, which is expressed normally ...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Takaoka N,Takayama T,Teratani T,Sugiyama T,Mugiya S,Ozono S

    更新日期:2011-07-19 00:00:00

  • Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5.

    abstract:BACKGROUND:Visceral leishmaniasis caused by members of the Leishmania donovani complex is often fatal in the absence of treatment. Research has been hampered by the lack of good laboratory models and tools for genetic manipulation. In this study, we have characterised a L. infantum line (JPCM5) that was isolated from a...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Denise H,Poot J,Jiménez M,Ambit A,Herrmann DC,Vermeulen AN,Coombs GH,Mottram JC

    更新日期:2006-11-13 00:00:00

  • New insights into SRY regulation through identification of 5' conserved sequences.

    abstract:BACKGROUND:SRY is the pivotal gene initiating male sex determination in most mammals, but how its expression is regulated is still not understood. In this study we derived novel SRY 5' flanking genomic sequence data from bovine and caprine genomic BAC clones. RESULTS:We identified four intervals of high homology upstr...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Ross DG,Bowles J,Koopman P,Lehnert S

    更新日期:2008-10-14 00:00:00

  • Identification of suitable reference genes for real-time quantitative PCR analysis of hydrogen peroxide-treated human umbilical vein endothelial cells.

    abstract:BACKGROUND:Oxidative stress can induce cell injury in vascular endothelial cells, which is the initial event in the development of atherosclerosis. Although quantitative real-time polymerase chain reaction (qRT-PCR) has been widely used in gene expression studies in oxidative stress injuries, using carefully validated ...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Li T,Diao H,Zhao L,Xing Y,Zhang J,Liu N,Yan Y,Tian X,Sun W,Liu B

    更新日期:2017-04-05 00:00:00

  • Selection of reference genes for normalisation of real-time RT-PCR in brain-stem death injury in Ovis aries.

    abstract:BACKGROUND:Heart and lung transplantation is frequently the only therapeutic option for patients with end stage cardio respiratory disease. Organ donation post brain stem death (BSD) is a pre-requisite, yet BSD itself causes such severe damage that many organs offered for donation are unusable, with lung being the orga...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Passmore M,Nataatmadja M,Fraser JF

    更新日期:2009-07-23 00:00:00

  • Small nucleolar RNA Sf-15 regulates proliferation and apoptosis of Spodoptera frugiperda Sf9 cells.

    abstract:BACKGROUND:Small nucleolar RNAs (snoRNAs) function in guiding 2'-O-methylation and pseudouridylation of ribosomal RNAs (rRNAs) and small nuclear RNAs (snRNAs). In recent years, more and more snoRNAs have been found to play novel roles in mRNA regulation, such as pre-mRNA splicing or RNA editing. In our previous study, ...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Wu B,Huang L,Qiu W,Liu X,Shen Y,Lu Y,Yang Z,Li X,Cui B,Xu S,Qiao H,Qiu R,Yao L,Kan Y,Li D

    更新日期:2019-04-11 00:00:00

  • SPRR2A enhances p53 deacetylation through HDAC1 and down regulates p21 promoter activity.

    abstract:BACKGROUND:Small proline rich protein (SPRR) 2A is one of 14 SPRR genes that encodes for a skin cross-linking protein, which confers structural integrity to the cornified keratinocyte cell envelope. New evidence, however, shows that SPRR2A is also a critical stress and wound repair modulator: it enables a variety of ba...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Mizuguchi Y,Specht S,Lunz JG 3rd,Isse K,Corbitt N,Takizawa T,Demetris AJ

    更新日期:2012-06-25 00:00:00