Abstract:
BACKGROUND:DsRed the red fluorescent protein (RFP) isolated from Discosoma sp. coral holds much promise as a genetically and spectrally distinct alternative to green fluorescent protein (GFP) for application in mice. Widespread use of DsRed has been hampered by several issues resulting in the inability to establish and maintain lines of red fluorescent protein expressing embryonic stem cells and mice. This has been attributed to the non-viability, or toxicity, of the protein, probably as a result of its obligate tetramerization. A mutagenesis approach directing the stepwise evolution of DsRed has produced mRFP1, the first true monomer. mRFP1 currently represents an attractive autofluorescent reporter for use in heterologous systems. RESULTS:We have used embryonic stem cell-mediated transgenesis to evaluate mRFP1 in embryonic stem cells and mice. We find that mRFP1 exhibits the most spatially homogenous expression when compared to the native (tetrameric) and variant dimeric forms of DsRed. High levels of mRFP1 expression do not affect cell morphology, developmental potential or viability and fertility of animals. High levels of widespread mRFP1 expression are maintained in a constitutive manner in embryonic stem cells in culture and in transgenic animals. We have used various optical imaging modalities to visualize mRFP1 expressing cells in culture, in embryos and adult mice. Moreover co-visualization of red, green and cyan fluorescent cells within a sample is easily achieved without the need for specialized methodologies, such as spectral deconvolution or linear unmixing. CONCLUSION:Fluorescent proteins with excitation and/or emission profiles in the red part of the visible spectrum represent distinct partners, or longer wavelength substitutes for GFP. Not only do DsRed-based RFPs provide a genetically and spectrally distinct addition to the available repertoire of autoflorescent proteins, but by virtue of their spectral properties they permit deeper tissue imaging. Our work in generating CAG::mRFP1 transgenic ES cells and mice demonstrates the developmental neutrality of mRFP1 in an organismal context. It paves the way for the use of DsRed-based monomeric RFPs in transgenic and gene targeted approaches which often necessitate spatially and/or temporally restricted reporter expression. Moreover animals of the CAG::mRFP1 transgenic strain serve as a source of RFP tagged tissue for the derivation of cell lines and explant, transplant and embryo chimera experiments.
journal_name
BMC Biotechnoljournal_title
BMC biotechnologyauthors
Long JZ,Lackan CS,Hadjantonakis AKdoi
10.1186/1472-6750-5-20keywords:
subject
Has Abstractpub_date
2005-07-04 00:00:00pages
20issn
1472-6750pii
1472-6750-5-20journal_volume
5pub_type
杂志文章abstract:BACKGROUND:Insect resistance in crops represents a main challenge for agriculture. Transgenic approaches based on proteins displaying insect resistance properties are widely used as efficient breeding strategies. To extend the spectrum of targeted pathogens and overtake the development of resistance, molecular evolutio...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0507-9
更新日期:2019-03-08 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:Over-expressed native or recombinant proteins are commonly used for industrial and pharmaceutical purposes, as well as for research. Proteins of interest need to be purified in sufficient quantity, quality and specific activity to justify their commercial price and eventual medical use. Proteome quality was ...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-016-0259-8
更新日期:2016-03-11 00:00:00
abstract:BACKGROUND:Fungal laccases are multicopper oxidases with huge applicability in different sectors. Here, we describe the development of a set of high-throughput colorimetric assays for screening laccase libraries in directed evolution studies. RESULTS:Firstly, we designed three colorimetric assays based on the oxidatio...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-13-90
更新日期:2013-10-26 00:00:00
abstract:BACKGROUND:A modification of a standard method of fluorescence in situ hybridisation (FISH) is described, by which a combination of several substrates and probes on single microscope slides enables more accurate comparisons of the distribution and abundance of chromosomal sequences and improves the relatively low throu...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-6-20
更新日期:2006-03-20 00:00:00
abstract:BACKGROUND:Serine hydroxymethyltransferase (SHMT) is the key enzyme in L-serine enzymatic production, suggesting the importance of obtaining a SHMT with high activity. RESULTS:Here, a novel SHMT gene, glyA, was obtained through degenerate oligonucleotide-primed PCR and encoded a novel SHMT with 54.3% similarity to the...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-014-0093-9
更新日期:2014-11-14 00:00:00
abstract:BACKGROUND:Heat-stable antifungal factor (HSAF) is a newly identified broad-spectrum antifungal antibiotic from the biocontrol agent Lysobacter enzymogenes and is regarded as a potential biological pesticide, due to its novel mode of action. However, the production level of HSAF is quite low, and little research has re...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-018-0478-2
更新日期:2018-10-24 00:00:00
abstract:BACKGROUND:Baculovirus, which has a width of 40 nm and a length of 250-300 nm, can display functional peptides, receptors and antigens on its surface by their fusion with a baculovirus envelop protein, GP64. In addition, some transmembrane proteins can be displayed without GP64 fusion, using the native transmembrane do...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-11-60
更新日期:2011-06-02 00:00:00
abstract:BACKGROUND:We have investigated the possibility and feasibility of producing the HPV-11 L1 major capsid protein in transgenic Arabidopsis thaliana ecotype Columbia and Nicotiana tabacum cv. Xanthi as potential sources for an inexpensive subunit vaccine. RESULTS:Transformation of plants was only achieved with the HPV-1...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-7-56
更新日期:2007-09-12 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: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:High recombinant protein productivity in mammalian cell lines is often associated with phenotypic changes in protein content, energy metabolism, and cell growth, but the key determinants that regulate productivity are still not clearly understood. The mammalian target of rapamycin (mTOR) signalling pathway h...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-14-15
更新日期:2014-02-17 00:00:00
abstract:BACKGROUND:Circular forms of viral genomic DNA are generated during infection of cells with retroviruses like HIV-1. Such circles are unable to replicate and are eventually lost as a result of cell division, lending support to the prevalent notion that episomal retroviral DNA forms are dead-end products of reverse tran...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-8-60
更新日期:2008-08-09 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:During a global influenza pandemic, the vaccine requirements of developing countries can surpass their supply capabilities, if these exist at all, compelling them to rely on developed countries for stocks that may not be available in time. There is thus a need for developing countries in general to produce t...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-12-14
更新日期:2012-04-26 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:Polysialic acid (polySia) is a carbohydrate modification of the neural cell adhesion molecule (NCAM), which is implicated in neural differentiation and plays an important role in tumor development and metastasis. Polysialylation of NCAM is mediated by two Golgi-resident polysialyltransferases (polyST) ST8Sia...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-017-0360-7
更新日期:2017-05-12 00:00:00
abstract:BACKGROUND:In a previous study, anthocyanin levels in potato plants were increased by manipulating genes connected with the flavonoid biosynthesis pathway. However, starch content and tuber yield were dramatically reduced in the transgenic plants, which over-expressed dihydroflavonol reductase (DFR). RESULTS:Transgeni...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-8-25
更新日期:2008-03-05 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 isothermal amplification of RNA in vitro has been used for the study of in vitro evolution of RNA. Although Qβ replicase has been traditionally used as an enzyme for this purpose, we planned to use norovirus replicase (NV3D(pol)) due to its structural simplicity in the scope of in vitro autonomous evolut...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-13-85
更新日期:2013-10-09 00:00:00
abstract:BACKGROUND:Advances in optical imaging modalities and the continued evolution of genetically-encoded fluorescent proteins are coming together to facilitate the study of cell behavior at high resolution in living organisms. As a result, imaging using autofluorescent protein reporters is gaining popularity in mouse trans...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-4-33
更新日期:2004-12-24 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: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 green fluorescent protein (GFP) has been regarded as a valuable tool and widely applied as a biomarker in medical applications and diagnostics. A cost-efficient upstream expression system and an inexpensive downstream purification process will meet the demands of the GFP protein with high-purity. RESULT...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0590-y
更新日期:2019-12-09 00:00:00
abstract:BACKGROUND:Aldehyde-deformylating oxygenase (ADO) is a key enzyme involved in the biosynthetic pathway of fatty alk(a/e)nes in cyanobacteria. However, cADO (cyanobacterial ADO) showed extreme low activity with the k cat value below 1 min-1, which would limit its application in biofuel production. To identify the activi...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-017-0351-8
更新日期:2017-03-16 00:00:00
abstract:BACKGROUND:More than a dozen of fungal immunomodulatory proteins (FIPs) have been identified to date, most of which are from Ganoderma species. However, little is known about the similarities and differences between different Ganoderma FIPs' bioactivities. In the current study, two FIP genes termed FIP-gap1 and FIP-gap...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-018-0488-0
更新日期:2018-12-14 00:00:00
abstract:BACKGROUND:Ligand-targeted approaches have proven successful in improving the therapeutic index of a number of drugs. We hypothesized that the specific targeting of TNF-alpha antagonists to inflamed tissues could increase drug efficacy and reduce side effects. RESULTS:Using uteroglobin (UG), a potent anti-inflammatory...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-11-104
更新日期:2011-11-10 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: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:There is an increasing need to understand cell-cell interactions for cell and tissue engineering purposes, such as optimizing cell sheet constructs, as well as for examining adhesion defect diseases. For cell-sheet engineering, one major obstacle to sheet function is that cell sheets in suspension are fragil...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-13-17
更新日期:2013-02-26 00:00:00