Abstract:
BACKGROUND:The use of small interfering RNA (siRNA) molecules in animals to achieve double-stranded RNA-mediated interference (RNAi) has recently emerged as a powerful method of sequence-specific gene knockdown. As DNA-based expression of short hairpin RNA (shRNA) for RNAi may offer some advantages over chemical and in vitro synthesised siRNA, a number of vectors for expression of shRNA have been developed. These often feature polymerase III (pol. III) promoters of either mouse or human origin. RESULTS:To develop a shRNA expression vector specifically for bovine RNAi applications, we identified and characterised a novel bovine U6 small nuclear RNA (snRNA) promoter from bovine sequence data. This promoter is the putative bovine homologue of the human U6-8 snRNA promoter, and features a number of functional sequence elements that are characteristic of these types of pol. III promoters. A PCR based cloning strategy was used to incorporate this promoter sequence into plasmid vectors along with shRNA sequences for RNAi. The promoter was then used to express shRNAs, which resulted in the efficient knockdown of an exogenous reporter gene and an endogenous bovine gene. CONCLUSION:We have mined data from the bovine genome sequencing project to identify a functional bovine U6 promoter and used the promoter sequence to construct a shRNA expression vector. The use of this native bovine promoter in shRNA expression is an important component of our future development of RNAi therapeutic and transgenic applications in bovine species.
journal_name
BMC Biotechnoljournal_title
BMC biotechnologyauthors
Lambeth LS,Moore RJ,Muralitharan M,Dalrymple BP,McWilliam S,Doran TJdoi
10.1186/1472-6750-5-13keywords:
subject
Has Abstractpub_date
2005-05-11 00:00:00pages
13issn
1472-6750pii
1472-6750-5-13journal_volume
5pub_type
杂志文章abstract:BACKGROUND:Cymbidium faberi is one of the oldest cultivars of oriental orchids, with an elegant flower fragrance. In order to investigate the molecular mechanism and the functions of related proteins in the methyl jasmonate (MeJA) signaling pathway, one of the main components of flower fragrance in C. faberi, yeast one...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-020-0599-2
更新日期:2020-01-16 00:00:00
abstract:BACKGROUND:Oleaginous yeast species are an alternative for the production of lipids or triacylglycerides (TAGs). These yeasts are usually non-pathogenic and able to store TAGs ranging from 20 % to 70 % of their cell mass depending on culture conditions. TAGs originating from oleaginous yeasts can be used as the so-call...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-016-0276-7
更新日期:2016-05-27 00:00:00
abstract:BACKGROUND:Expression vector engineering technology is one of the most convenient and timely method for cell line development to meet the rising demand of novel production cell line with high productivity. Destabilization of dihydrofolate reductase (dhfr) selection marker by addition of AU-rich elements and murine orni...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-015-0145-9
更新日期:2015-06-02 00:00:00
abstract:BACKGROUND:The identification of known mutations in a cell population is important for clinical applications and basic cancer research. In this work an immobilized form of the polymerase chain reaction, referred to as polony technology, was used to detect mutations as well as gene deletions, resulting in loss of hetero...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-3-11
更新日期:2003-07-23 00:00:00
abstract:BACKGROUND:CRISPR/Cas9 is widely used for precise genetic editing in various organisms. CRISPR/Cas9 editing may in many plants be hampered by the presence of complex and high ploidy genomes and inefficient or poorly controlled delivery of the CRISPR/Cas9 components to gamete cells or cells with regenerative potential. ...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0530-x
更新日期:2019-06-17 00:00:00
abstract:BACKGROUND:Hydrophobic polystyrene is the most common material for solid phase immunoassay. Proteins are immobilized on polystyrene by passive adsorption, which often causes considerable denaturation. Biological macromolecules were found to better retain their functional activity when immobilized on hydrophilic materia...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-1-4
更新日期:2001-01-01 00:00:00
abstract:BACKGROUND:The ability of cytosine deaminase (CD) to convert the antifungal agent 5-fluorocytosine (5-FC) into one of the most potent and largely used anticancer compound such as 5-fluorouracil (5-FU) raised considerable interest in this enzyme to model gene or antibody - directed enzyme-prodrug therapy (GDEPT/ADEPT) a...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-8-68
更新日期:2008-09-10 00:00:00
abstract:BACKGROUND:Multi-copy gene integration into microbial genomes is a conventional tool for obtaining improved gene expression. For Penicillium chrysogenum, the fungal producer of the beta-lactam antibiotic penicillin, many production strains carry multiple copies of the penicillin biosynthesis gene cluster. This discover...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-017-0335-8
更新日期:2017-02-16 00:00:00
abstract:BACKGROUND:Heparan sulfate glycosaminoglycans are diverse components of certain proteoglycans and are known to interact with growth factors as a co-receptor necessary to induce signalling and growth factor activity. In this report we characterize heterogeneously glycosylated recombinant human perlecan domain 1 (HSPG2 a...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-10-66
更新日期:2010-09-09 00:00:00
abstract:BACKGROUND:Interest in using plants for production of recombinant proteins such as monoclonal antibodies is growing, but proteolytic degradation, leading to a loss of functionality and complications in downstream purification, is still a serious problem. RESULTS:In this study, we investigated the dynamics of the assem...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-11-128
更新日期:2011-12-30 00:00:00
abstract:BACKGROUND:Protein cross-coupling reactions demand high yields, especially if the products are intended for bioanalytics, like enzyme-linked immunosorbent assays. Amongst other factors, the coupling yield depends on the concentration of the proteins being used for coupling. Protein supercharging of enzymes can increase...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-014-0088-6
更新日期:2014-10-18 00:00:00
abstract:BACKGROUND:The CRISPR/Cas9 system is increasingly used for gene inactivation in mouse zygotes, but homology-directed mutagenesis and use of inbred embryos are less established. In particular, Rosa26 knock-in alleles for the insertion of transgenes in a genomic 'safe harbor' site, have not been produced. Here we applied...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-016-0234-4
更新日期:2016-01-16 00:00:00
abstract:BACKGROUND:Many growth factors, such as bone morphogenetic protein (BMP)-2, have been shown to interact with polymers of sulfated disacharrides known as heparan sulfate (HS) glycosaminoglycans (GAGs), which are found on matrix and cell-surface proteoglycans throughout the body. HS GAGs, and some more highly sulfated fo...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-12-60
更新日期:2012-09-11 00:00:00
abstract:BACKGROUND:Ultrasound and microbubbles (USMB) have been shown to enhance the intracellular uptake of molecules, generally thought to occur as a result of sonoporation. The underlying mechanism associated with USMB-enhanced intracellular uptake such as membrane disruption and endocytosis may also be associated with USMB...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-017-0364-3
更新日期:2017-05-18 00:00:00
abstract:BACKGROUND:Rapid, robust and reversible induction of transgene expression would significantly facilitate cancer gene therapy as well as allow the in vivo functional study of newly discovered genes in tumor formation and progression. The popularity of the ecdysone inducible gene switch system has led us to investigate w...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-1-11
更新日期:2001-01-01 00:00:00
abstract:BACKGROUND:Loop mediated isothermal amplification of nucleic acid templates is a rapid, sensitive and specific method suitable for molecular diagnostics. However the complexity of primer design and the number of primers involved can lead to false positives from non-specific primer interactions. Standard methods of LAMP...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0549-z
更新日期:2019-08-01 00:00:00
abstract:BACKGROUND:The use of highly reproducible and spatiallyhomogeneous thin film matrices permits automated microscopy and quantitative determination of the response of hundreds of cells in a population. Using thin films of extracellular matrix proteins, we have quantified, on a cell-by-cell basis, phenotypic parameters of...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-6-14
更新日期:2006-03-06 00:00:00
abstract:BACKGROUND:In the previous study, the cellulolytic Escherichia coli ZH-4 isolated from bovine rumen was found to show extracellular cellulase activity and could degrade cellulose in the culture. The goal of this work was to identify and characterize the secreted cellulase of E. coli ZH-4. It will be helpful to re-under...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0556-0
更新日期:2019-08-27 00:00:00
abstract:BACKGROUND:The therapeutic use of α-amidated peptides (e.g. calcitonin, glucagon-like peptide) has increased dramatically, but there are major impediments to wider use of such peptides. Larger peptides are expensive to synthesize, and short plasma half-lives frequently limit the clinical circumstances in which the pept...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-015-0173-5
更新日期:2015-06-27 00:00:00
abstract:BACKGROUND:Dengue is a public health problem of global significance for which there is neither an effective antiviral therapy nor a preventive vaccine. It is a mosquito-borne viral disease, caused by dengue (DEN) viruses, which are members of the Flaviviridae family. There are four closely related serotypes, DEN-1, DEN...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-7-10
更新日期:2007-02-15 00:00:00
abstract:BACKGROUND:Nanoparticles appear to be promising devices for application in the agriculture and food industries, but information regarding the response of plants to contact with nano-devices is scarce. Toxic effects may be imposed depending on the type and concentration of nanoparticle as well as time of exposure. A num...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-13-111
更新日期:2013-12-20 00:00:00
abstract:BACKGROUND:Understanding and manipulating gene function in physiological conditions is a major objective for both fundamental and applied research. In contrast to other experimental settings, which use either purely genetic or gene delivery (viral or non-viral) strategies, we report here a strategy based on direct prot...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-9-40
更新日期:2009-04-24 00:00:00
abstract:BACKGROUND:Acetyl-xylan esterase (AXE, EC 3.1.1.72) hydrolyses acetate group from the linear chain of xylopyranose residues bound by β-1,4-linkage. The enzyme finds commercial applications in bio-bleaching of wood pulp, treating animal feed to increase digestibility, processing food to increase clarification and conver...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-11-52
更新日期:2011-05-16 00:00:00
abstract:BACKGROUND:Prostate cancer is the most-diagnosed non-skin cancer among males in the US, and the second leading cause of cancer-related death. Current methods of treatment and diagnosis are not specific for the disease. This work identified an antibody fragment that binds selectively to a molecule on the surface of andr...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-14-81
更新日期:2014-09-03 00:00:00
abstract:BACKGROUND:Carboxyl-terminal protease (CtpA) plays essential functions in posttranslational protein processing in prokaryotic and eukaryotic cells. To date, only a few bacterial ctpA genes have been characterized. Here we cloned and characterized a novel CtpA. The encoding gene, ctpAp (ctpA of Paenibacillus lautus), wa...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-13-89
更新日期:2013-10-25 00:00:00
abstract:BACKGROUND:Engineered proteins, with non-immunoglobulin scaffolds, have become an important alternative to antibodies in many biotechnical and therapeutic applications. When compared to antibodies, tailored proteins may provide advantageous properties such as a smaller size or a more stable structure. RESULTS:Avidin i...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-11-64
更新日期:2011-06-09 00:00:00
abstract:BACKGROUND:Despite positive reports on the efficacy of stem cell therapy for the treatment of cardiovascular disease, the nature of stem cell homing to ischemic tissues remains elusive. RESULTS:We used a mouse model of peripheral tissue ischemia to study the survival and homing capacity of dual reporter gene (eGFP/Luc...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-12-93
更新日期:2012-12-03 00:00:00
abstract:BACKGROUND:Clostridial co-culture containing cellulolytic and solventogenic species is a potential consolidated bioprocessing (CBP) approach for producing biochemicals and biofuels from cellulosic biomass. It has been demonstrated that the rate of cellulose utilization in the co-culture of Clostridium acetobutylicum an...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-13-95
更新日期:2013-11-04 00:00:00
abstract:BACKGROUND:Each year, influenza is responsible for hundreds of thousand cases of illness and deaths worldwide. Due to the virus' fast mutation rate, the World Health Organization (WHO) is constantly on alert to rapidly respond to emerging pandemic strains. Although anti-viral therapies exist, the most proficient way to...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-015-0152-x
更新日期:2015-05-16 00:00:00
abstract:BACKGROUND:Vascular endothelial growth factor (VEGF) is a key angiogenic factors. It plays an important role in both physiologic and pathologic angiogenesis and increases permeability across the vessels. Using antibody phage display technology, we obtained a novel anti-VEGFA IgG, named as FD006. In this study, the phar...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-14-17
更新日期:2014-02-27 00:00:00