Abstract:
BACKGROUND:In spite of a great number of monomeric fluorescent proteins developed in the recent years, the reported fluorescent protein-based FRET pairs are still characterized by a number of disadvantageous features, complicating their use as reporters in cell biology and for high-throughput cell-based screenings. RESULTS:Here we screened some of the recently developed monomeric protein pairs to find the optimal combination, which would provide high dynamic range FRET changes, along with high pH- and photo-stability, fast maturation and bright fluorescence, and reliable detection in any fluorescent imaging system. Among generated FRET pairs, we have selected TagGFP-TagRFP, combining all the mentioned desirable characteristics. On the basis of this highly efficient FRET pair, we have generated a bright, high contrast, pH- and photo-stable apoptosis reporter, named CaspeR3 (Caspase 3 Reporter). CONCLUSION:The combined advantages suggest that the TagGFP-TagRFP is one of the most efficient green/red couples available to date for FRET/FLIM analyses to monitor interaction of proteins of interest in living cells and to generate FRET-based sensors for various applications. CaspeR3 provides reliable detection of apoptosis, and should become a popular tool both for cell biology studies and high throughput screening assays.
journal_name
BMC Biotechnoljournal_title
BMC biotechnologyauthors
Shcherbo D,Souslova EA,Goedhart J,Chepurnykh TV,Gaintzeva A,Shemiakina II,Gadella TW,Lukyanov S,Chudakov DMdoi
10.1186/1472-6750-9-24subject
Has Abstractpub_date
2009-03-25 00:00:00pages
24issn
1472-6750pii
1472-6750-9-24journal_volume
9pub_type
杂志文章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: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:Arachidonic acid (ARA, C20:4, n-6), which belongs to the omega-6 series of polyunsaturated fatty acids and has a variety of biological activities, is commercially produced in Mortierella alpina. Dissolved oxygen or oxygen utilization efficiency is a critical factor for Mortierella alpina growth and arachidon...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-017-0388-8
更新日期:2017-08-30 00:00:00
abstract:BACKGROUND:Proteins in inclusion bodies (IBs) present native-like secondary structures. However, chaotropic agents at denaturing concentrations, which are widely used for IB solubilization and subsequent refolding, unfold these secondary structures. Removal of the chaotropes frequently causes reaggregation and poor rec...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-018-0486-2
更新日期:2018-12-12 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:There is an imperative necessity for alternative sources of energy able to reduce the world dependence of fossil oil. One of the most successful options is ethanol obtained mainly from sugarcane and corn fermentation. The foremost residue from sugarcane industry is the bagasse, a rich lignocellulosic raw mat...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-13-94
更新日期:2013-10-31 00:00:00
abstract:BACKGROUND:Domesticated einkorn (Triticum monococcum L.) is one of the oldest cultivated cereal crops in the world. Its small genome size (~ 5.7 GB), low ploidy (2n = 2x = 14, AmAm) and high genetic polymorphism make this species very attractive for use as a diploid model for understanding the genomics and proteomics o...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-018-0477-3
更新日期:2018-10-23 00:00:00
abstract:BACKGROUND:The presence of coliform bacteria is routinely assessed to establish the microbiological safety of water supplies and raw or processed foods. Coliforms are a group of lactose-fermenting Enterobacteriaceae, which most likely acquired the lacZ gene by horizontal transfer and therefore constitute a polyphyletic...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-015-0168-2
更新日期:2015-06-04 00:00:00
abstract:BACKGROUND:Interleukin-17 (IL-17), the characteristic cytokine secreted by T helper 17 lymphocytes (Th17 cells), plays a pivotal role in host defense and many inflammatory or autoimmune diseases. The aim of this study was to obtain purified protein caprine IL-17A (cIL-17A) as an antigen for preparing an IL-17A-specific...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0543-5
更新日期:2019-07-17 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:Spinal cord injury (SCI) remains a huge medical problem nowadays as there is no hospital providing the versatile strategies for repairing central nervous system and restoring function. Herein, we focused on PC-12 cells as an important research tool and studied the potential role of resveratrol (RSV) in infla...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0502-1
更新日期:2019-01-28 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: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:Microarray technology has been used extensively over the past 10 years for assessing gene expression, and has facilitated precise genetic profiling of everything from tumors to small molecule drugs. By contrast, arraying cell membranes in a manner which preserves their ability to mediate biochemical processe...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-5-18
更新日期:2005-06-16 00:00:00
abstract:BACKGROUND:In vivo use of monoclonal antibodies has become routine clinical practice in the treatment of human cancer. CD38 is an attractive target, because it has double roles, as a receptor and an ectoenzyme. Daratumumab, an anti-CD38 antibody, is currently in the clinical trials for multiple myeloma. RESULTS:Here w...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0524-8
更新日期:2019-05-22 00:00:00
abstract:BACKGROUND:Shark heavy chain antibody, also called new antigen receptor (NAR), consists of one single Variable domain (VH), containing only two complementarity-determining regions (CDRs). The antigen binding affinity and specificity are mainly determined by these two CDRs. The good solubility, excellent thermal stabili...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-7-78
更新日期:2007-11-19 00:00:00
abstract:BACKGROUND:In the search for generic expression strategies for mammalian protein families several bacterial expression vectors were examined for their ability to promote high yields of soluble protein. Proteins studied included cell surface receptors (Ephrins and Eph receptors, CD44), kinases (EGFR-cytoplasmic domain, ...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-4-32
更新日期:2004-12-14 00:00:00
abstract:BACKGROUND:Protein glycosylation is of fundamental importance in many biological systems. The discovery of N-glycosylation in bacteria and the functional expression of the N-oligosaccharyltransferase PglB of Campylobacter jejuni in Escherichia coli enabled the production of engineered glycoproteins and the study of the...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-12-67
更新日期:2012-09-24 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:Construction of high quality cDNA libraries from the usually low amounts of eukaryotic mRNA extracted from environmental samples is essential in functional metatranscriptomics for the selection of functional, full-length genes encoding proteins of interest. Many of the inserts in libraries constructed by sta...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-14-80
更新日期:2014-09-03 00:00:00
abstract:BACKGROUND:One of the most commonly used vectors for gene therapy is the adenoviral vector; its ability to tightly regulate transgene expression is critical for optimizing therapeutic outcomes. The tetracycline-regulated system (especially the Tet-On system) for gene expression is one of the most valuable tools for con...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-015-0121-4
更新日期:2015-02-13 00:00:00
abstract:BACKGROUND:Bacterial degradation/transformation of steroids is widely investigated to create biotechnologically relevant strains for industrial application. The strain of Nocardioides simplex VKM Ac-2033D is well known mainly for its superior 3-ketosteroid Δ1-dehydrogenase activity towards various 3-oxosteroids and oth...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-021-00668-9
更新日期:2021-01-13 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
abstract:BACKGROUND:Plants are increasingly being examined as alternative recombinant protein expression systems. Recombinant protein expression levels in plants from Tobacco mosaic virus (TMV)-based vectors are much higher than those possible from plant promoters. However the common TMV expression vectors are costly, and at ti...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-7-52
更新日期:2007-08-27 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:The insect cell line is a critical component in the production of recombinant proteins in the baculovirus expression system and new cell lines hold the promise of increasing both quantity and quality of protein production. RESULTS:Seventy cell lines were established by single-cell cloning from a primary cul...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-10-50
更新日期:2010-07-05 00:00:00
abstract:BACKGROUND:Cost-effective production of industrially important enzymes is a key for their successful exploitation on industrial scale. Keeping in view the extensive industrial applications of lignin peroxidase (LiP), this study was performed to purify and characterize the LiP from an indigenous strain of Trametes versi...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-12-46
更新日期:2012-08-03 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: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