Abstract:
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.
journal_name
BMC Mol Bioljournal_title
BMC molecular biologyauthors
Magnani E,Bartling L,Hake Sdoi
10.1186/1471-2199-7-46subject
Has Abstractpub_date
2006-12-06 00:00:00pages
46issn
1471-2199pii
1471-2199-7-46journal_volume
7pub_type
杂志文章abstract:BACKGROUND:Germline mutations in the tumor suppressor PTEN predispose human beings to breast cancer, and genetic and epigenetic alterations of PTEN are also detected in sporadic human breast cancer. Germline Pten mutations in mice lead to the development of a variety of tumors, but mammary carcinomas are infrequently f...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-2-2
更新日期:2001-01-01 00:00:00
abstract:BACKGROUND:CCAAT/Enhancer Binding Proteindelta (C/EBPdelta) is a member of the highly conserved C/EBP family of leucine zipper (bZIP) proteins. C/EBPdelta is highly expressed in G0 growth arrested mammary epithelial cells (MECs) and "loss of function" alterations in C/EBPdelta have been associated with impaired contact...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-9-83
更新日期:2008-10-01 00:00:00
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
doi:10.1186/1471-2199-14-21
更新日期:2013-09-12 00:00:00
abstract:BACKGROUND:The human adenovirus type 5 early region 1A (E1A) gene encodes proteins that are potent regulators of transcription. E1A does not bind DNA directly, but is recruited to target promoters by the interaction with sequence specific DNA binding proteins. In mammalian systems, E1A has been shown to contain two reg...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-10-32
更新日期:2009-04-17 00:00:00
abstract:BACKGROUND:Cdc23/Mcm10 is required for the initiation and elongation steps of DNA replication but its biochemical function is unclear. Here, we probe its function using a novel approach in fission yeast, involving Cdc23 cleavage by the TEV protease. RESULTS:Insertion of a TEV protease cleavage site into Cdc23 allows i...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-6-13
更新日期:2005-06-07 00:00:00
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
doi:10.1186/1471-2199-10-72
更新日期:2009-07-23 00:00:00
abstract:BACKGROUND:Conserved two-component system ColRS of Pseudomonas genus has been implicated in several unrelated phenotypes. For instance, deficiency of P. putida ColRS system results in lowered phenol tolerance, hindrance of transposition of Tn4652 and lysis of a subpopulation of glucose-grown bacteria. In order to disco...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-10-46
更新日期:2009-05-16 00:00:00
abstract:BACKGROUND:The rapid increase in whole genome fungal sequence information allows large scale functional analyses of target genes. Efficient transformation methods to obtain site-directed gene replacement, targeted over-expression by promoter replacement, in-frame epitope tagging or fusion of coding sequences with fluor...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-9-70
更新日期:2008-08-01 00:00:00
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
doi:10.1186/1471-2199-13-20
更新日期:2012-06-25 00:00:00
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
doi:10.1186/s12867-019-0120-4
更新日期:2019-01-15 00:00:00
abstract:BACKGROUND:In spite of large intergenic spaces in plant and animal genomes, 7% to 30% of genes in the genomes encode overlapping cis-natural antisense transcripts (cis-NATs). The widespread occurrence of cis-NATs suggests an evolutionary advantage for this type of genomic arrangement. Experimental evidence for the regu...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-9-6
更新日期:2008-01-14 00:00:00
abstract:BACKGROUND:Pseudo-nitzschia multiseries Hasle (Hasle) (Ps-n) is distinctive among the ecologically important marine diatoms because it produces the neurotoxin domoic acid. Although the biology of Ps-n has been investigated intensely, the characterization of the genes and biochemical pathways leading to domoic acid bios...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-14-25
更新日期:2013-11-01 00:00:00
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
doi:10.1186/1471-2199-15-10
更新日期:2014-05-25 00:00:00
abstract:BACKGROUND:Retinal ganglion cells (RGCs) are preferentially lost in glaucoma or optic neuritis. In the present study, we investigated the protective effect of mircoRNA 100 (miR-100) against oxidative stress induced apoptosis in RGC-5 cells. RESULTS:Rat RGC-5 cells were cultured in plates and H2O2 was added to induce o...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/s12867-014-0025-1
更新日期:2014-11-18 00:00:00
abstract:BACKGROUND:In budding yeast, the replication checkpoint slows progress through S phase by inhibiting replication origin firing. In mammals, the replication checkpoint inhibits both origin firing and replication fork movement. To find out which strategy is employed in the fission yeast, Schizosaccharomyces pombe, we use...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-8-112
更新日期:2007-12-19 00:00:00
abstract:BACKGROUND:Transient Receptor Potential Canonical 1 (TRPC1) is a widely-expressed mammalian cationic channel with functional effects that include stimulation of cardiovascular remodelling. The initial aim of this study was to investigate variation in TRPC1-encoding gene transcripts. RESULTS:Extensive TRPC1 transcript ...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-12-30
更新日期:2011-07-12 00:00:00
abstract:BACKGROUND:Single-stranded DNA-binding proteins are essential cellular components required for the protection, metabolism and processing of single-stranded DNA. Human single-stranded DNA-binding protein 1 (hSSB1) is one such protein, with described roles in genome stability maintenance and transcriptional regulation. A...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/s12867-016-0077-5
更新日期:2016-12-09 00:00:00
abstract:BACKGROUND:When estimating relative transcript abundances by quantitative real-time PCR (Q-PCR) we found that the results can vary dramatically depending on the method chosen for data analysis. RESULTS:Analyses of Q-PCR results from a salmon louse starvation experiment show that, even with apparently good raw data, di...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-6-10
更新日期:2005-04-26 00:00:00
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
doi:10.1186/1471-2199-2-12
更新日期:2001-01-01 00:00:00
abstract:BACKGROUND:Macrophages (Mtheta) play a central role in the innate immune response and in the pathology of chronic inflammatory diseases. Macrophages treated with Th2-type cytokines such as Interleukin-4 (IL-4) and Interleukin-13 (IL-13) exhibit an altered phenotype and such alternatively activated macrophages are impor...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-7-45
更新日期:2006-11-29 00:00:00
abstract:BACKGROUND:ATPase/Helicases and nucleases play important roles in homologous recombination repair (HRR). Many of the mechanistic details relating to these enzymes and their function in this fundamental and complicated DNA repair process remain poorly understood in archaea. Here we employed Sulfolobus islandicus, a hype...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/s12867-015-0030-z
更新日期:2015-02-14 00:00:00
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
doi:10.1186/1471-2199-9-96
更新日期:2008-10-30 00:00:00
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
doi:10.1186/1471-2199-9-50
更新日期:2008-05-19 00:00:00
abstract:BACKGROUND:Japanese encephalitis virus (JEV) NS5 is a viral nonstructural protein that carries both methyltransferase and RNA-dependent RNA polymerase (RdRp) domains. It is a key component of the viral RNA replicase complex that presumably includes other viral nonstructural and cellular proteins. The biochemical proper...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-8-59
更新日期:2007-07-11 00:00:00
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
doi:10.1186/s12867-019-0128-9
更新日期:2019-04-11 00:00:00
abstract:BACKGROUND:Cyclic AMP-dependent protein kinase (PKA) plays a central role in regulation of energy metabolism. Upon stimulation of testicular Sertoli cells by follicle stimulating hormone (FSH), glycolysis is activated to increase the production of nutrients for the germ cells, and a new regulatory subunit of cAMP-depen...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-3-10
更新日期:2002-06-21 00:00:00
abstract:BACKGROUND:Retrotransposition is an important evolutionary force for the creation of new and potentially functional intronless genes which are collectively called retrogenes. Many retrogenes are expressed in the testis and the gene products have been shown to actively participate in spermatogenesis and other unique fun...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-10-74
更新日期:2009-07-25 00:00:00
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
doi:10.1186/1471-2199-10-55
更新日期:2009-06-05 00:00:00
abstract:BACKGROUND:It is not clear how foreign DNA molecules insert into the host genome. Recently, we have produced transgenic mice to investigate the role of the fad2 gene in the conversion of oleic acid to linoleic acid. Here we describe an integration mechanism of fad2 transgene by homologous illegitimate random integratio...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-11-58
更新日期:2010-08-13 00:00:00
abstract:BACKGROUND:The majority of protein isoforms arise from alternative splicing of the encoding primary RNA transcripts. To understand the significance of single splicing events, reliable techniques are needed to determine their incidence. However, existing methods are labour-intensive, error-prone or of limited use. RESU...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/s12867-016-0060-1
更新日期:2016-03-15 00:00:00