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 interest (tester-tester) are filled-in with alpha-thio-deoxynucleotides. Unprotected duplexes, as well as the single-stranded DNA fragments, are degraded using ExoIII and Mung Bean Nuclease, prior to PCR subtraction, resulting in less complex difference products. We illustrate this modification by the cloning of a new gene which is differentially expressed in normal, reelin and Dab1 mutant mice and is a candidate member of the Reelin signalling pathway involved in brain development. CONCLUSION:We propose a modification of cDNA-RDA that may reduce the complexity of the post-hybridization mix and thus facilitate the amplification of differentially expressed products.
journal_name
BMC Mol Bioljournal_title
BMC molecular biologyauthors
Kuvbachieva AA,Goffinet AMdoi
10.1186/1471-2199-3-6keywords:
subject
Has Abstractpub_date
2002-04-24 00:00:00pages
6issn
1471-2199journal_volume
3pub_type
杂志文章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: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: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
doi:10.1186/1471-2199-7-23
更新日期:2006-07-18 00:00:00
abstract:BACKGROUND:Although staphylokianse (SAK) is among the most promising blood dissolving agents, it is far from ideal. It is interesting to hypothesize that the clot lysis efficacy of SAK can be enhanced with direct active platelet binding ability, and at the same time the rethrombosis complication after successful recana...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-8-88
更新日期:2007-10-07 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: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
doi:10.1186/1471-2199-12-31
更新日期:2011-07-19 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: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:Bothrops colombiensis is a highly dangerous pit viper and responsible for over 70% of snakebites in Venezuela. Although the composition in B. colombiensis venom has been identified using a proteome analysis, the venom gland transcriptome is currently lacking. RESULTS:We constructed a cDNA library from the v...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/s12867-016-0059-7
更新日期:2016-03-05 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: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: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: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
doi:10.1186/s12867-016-0057-9
更新日期:2016-02-09 00:00:00
abstract:BACKGROUND:Nucleosomes are nucleoproteic complexes, formed of eight histone molecules and DNA, and they are responsible for the compaction of the eukaryotic genome. Their presence on DNA influences many cellular processes, such as transcription, DNA replication, and DNA repair. The evolutionarily conserved histone vari...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-10-18
更新日期:2009-03-04 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: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: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: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: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: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: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:The Y chromosome of mammals is particularly prone to accumulate genes related to male fertility. However, the high rate of molecular evolution on this chromosome predicts reduced power to the across-species comparative approach in identifying male-specific genes that are essential for sperm production in hum...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-11-33
更新日期:2010-05-07 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: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: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: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: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
doi:10.1186/1471-2199-4-5
更新日期:2003-04-28 00:00:00
abstract:BACKGROUND:We reported previously that the cardiac troponin I (cTnI) promoter drives cardiac-specific expression of reporter genes in cardiac muscle cells and in transgenic mice, and that disruption of GATA elements inactivates the cTnI promoter in cultured cardiomyocytes. We have now examined the role of cTnI promoter...
journal_title:BMC molecular biology
pub_type: 杂志文章
doi:10.1186/1471-2199-8-78
更新日期:2007-09-17 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: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
doi:10.1186/1471-2199-9-85
更新日期:2008-10-14 00:00:00