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:Physiological long term adaptation of pancreatic beta cells is driven by stimuli such as glucose and incretin hormones acting via cAMP (e.g. GLP-1) and involves regulated gene expression. Several rapidly inducible immediate-early genes (IEGs) have been identified in beta cells. Many of these IEGs code for tr...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-8-54
更新日期:2007-06-22 00:00:00
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
doi:10.1186/s12867-017-0086-z
更新日期:2017-04-05 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:Previous studies suggest that human RAD9 (hRad9), encoding a DNA damage checkpoint molecule, which is frequently amplified in epithelial tumor cells of breast, lung, head and neck cancer, participates in regulation of the tumor suppressor p53-dependent transactivation of pro-survival P21WAF1. This study exam...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-8-37
更新日期:2007-05-21 00:00:00
abstract:BACKGROUND:Carbonic anhydrases (CAs) are found in many organisms, in which they contribute to several important biological processes. The vertebrate alpha-CA family consists of 16 subfamilies, three of which (VIII, X and XI) consist of acatalytic proteins. These are named carbonic anhydrase related proteins (CARPs), an...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-11-25
更新日期:2010-03-31 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:Although much is known about molecular mechanisms that prevent re-initiation of DNA replication on newly replicated DNA during a single cell cycle, knowledge is sparse regarding the regions that are most susceptible to re-replication when those mechanisms are bypassed and regarding the extents to which check...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-8-119
更新日期:2007-12-21 00:00:00
abstract:BACKGROUND:ALL1-fused from chromosome 1q (AF1q), originally considered as an oncogenic factor, has been implicated in neuronal development; however, its upstream regulatory mechanisms in neural system remained elusive. RESULTS:Our study showed that REST (RE1 silencing transcription factor), a key transcription factor ...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/s12867-015-0043-7
更新日期:2015-09-05 00:00:00
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
doi:10.1186/1471-2199-8-27
更新日期:2007-04-12 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: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: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
doi:10.1186/1471-2199-8-96
更新日期:2007-10-25 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
abstract:BACKGROUND:Cryptochromes (CRYs) are a class of flavoprotein blue-light signaling receptors found in plants and animals, and they control plant development and the entrainment of circadian rhythms. They also act as integral parts of the central circadian oscillator in humans and other animals. In mammals, the CLOCK-BMAL...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-11-69
更新日期:2010-09-14 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: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: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: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
doi:10.1186/1471-2199-5-6
更新日期:2004-07-01 00:00:00
abstract:BACKGROUND:The selection of stable and suitable reference genes for real-time quantitative PCR (RT-qPCR) is a crucial prerequisite for reliable gene expression analysis under different experimental conditions. The present study aimed to identify reference genes as internal controls for gene expression studies by RT-qPC...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-13-22
更新日期:2012-06-29 00:00:00
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
doi:10.1186/1471-2199-5-14
更新日期:2004-08-26 00:00:00
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
doi:10.1186/1471-2199-11-64
更新日期:2010-08-27 00:00:00
abstract:BACKGROUND:hAT elements and V(D)J recombination may have evolved from a common ancestral transposable element system. Extrachromosomal, circular forms of transposable elements (referred to here as episomal forms) have been reported yet their biological significance remains unknown. V(D)J signal joints, which resemble e...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-10-108
更新日期:2009-12-14 00:00:00
abstract:BACKGROUND:Many studies of the eukaryotic transcription mechanism and its regulation rely on in vitro assays. Conventional RNA polymerase II transcription assays are based on radioactive labelling of the newly synthesized RNA. Due to the inefficient in vitro transcription, the detection of the RNA involving purificatio...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-15-7
更新日期:2014-04-03 00:00:00
abstract:BACKGROUND:Members of the Sox gene family isolated from both vertebrates and invertebrates have been proved to participate in a wide variety of developmental processes, including sex determination and differentiation. Among these members, Sox30 had been considered to exist only in mammals since its discovery, and its e...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-11-98
更新日期:2010-12-11 00:00:00
abstract:BACKGROUND:Senescence marker protein-30 (SMP30), whose expression declines during aging in rat liver, has been proposed as an important aging marker. Besides apoptosis, SMP30 also protects cells against various other injuries by enhancement of membrane calcium-pump activity. The mechanism of this differential gene expr...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-9-43
更新日期:2008-04-29 00:00:00
abstract:BACKGROUND:Core promoters are cis-regulatory modules to which bind the basal transcriptional machinery and which participate in the regulation of transcription initiation. Although core promoters have not been extensively investigated through functional assays in a chromosomal context, the available data suggested that...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-12-32
更新日期:2011-08-01 00:00:00
abstract:BACKGROUND:Core canonical histones are required in the S phase of the cell cycle to pack newly synthetized DNA, therefore the expression of their genes is highly activated during DNA replication. In mammalian cells, this increment is achieved by both enhanced transcription and 3' end processing. In this paper, we descr...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/s12867-018-0110-y
更新日期:2018-07-27 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: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:Among the three functions of DNA, mammalian replication and transcription can be subject to epigenetic imprinting specified by the parental origin of chromosomes, and although there is suggestive indication that this is also true for meiotic recombination, no definitive evidence has yet been reported. RESUL...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-10-43
更新日期:2009-05-13 00:00:00