Abstract：
:One of the major constraints to performing large-scale transfections of cultured mammalian cells for the transient expression of recombinant proteins is the production of large quantities of purified plasmid DNA. In this report partially purified plasmid DNA was prepared by a method that combines alkaline lysis of E. coli with standard precipitation techniques. The efficiency of calcium phosphate-DNA co-precipitate formation with crude DNA was similar to that observed for pure DNA, but precipitate formed with crude DNA also contained RNA. The transfection of adherent and suspension-adapted HEK293-EBNA cells with partially purified pEGFPN1 resulted in levels of transient GFP expression equivalent to those achieved with pure DNA. In addition, the co-transfection of 1-200 ml cultures of suspension-adapted HEK293-EBNA cells with two different plasmids encoding the heavy and light chain genes of anti-human RhD IgG1, respectively, yielded similar IgG titers with pure and partially purified plasmid DNA. Finally, it was observed that suspension-adapted cells were more tolerant to the presence of RNA in the plasmid preparations than were adherent cells. These findings are relevant to the field of DNA transfection, including applications ranging from high-throughput screening to large-scale transient protein expression.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Wright JL,Jordan M,Wurm FMdoi
10.1016/s0168-1656(03)00032-4subject
Has Abstractpub_date
2003-05-08 00:00:00pages
211-21issue
3eissn
0168-1656issn
1873-4863pii
S0168165603000324journal_volume
102pub_type
杂志文章abstract：:The action of three glycoside hydrolase family GH51 and one GH54 α-arabinofuranosidases (ABFs) was studied on polymeric arabinoxylan and arabinoxylooligosaccharides. The substrates covered all possible arabinofuranosyl (Araf) substituents, i.e., terminal and internal α(1→2) and α(1→3)-linked Araf monosubstitutions and...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2016.04.050
更新日期：2016-07-10 00:00:00
abstract：:Based on the importance of heat shock proteins (HSPs) in diseases such as cancer, Alzheimer's disease or malaria, inhibitors of these chaperons are needed. Today's state-of-the-art techniques to identify HSP inhibitors are performed in microplate format, requiring large amounts of proteins and potential inhibitors. In...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.03.006
更新日期：2014-06-20 00:00:00
abstract：:A simple and an efficient oligonucleotide array was developed to identify common severe determinants of alpha (alpha) thalassemia. A total of 14 probes were designed to detect the most frequently three deletions (-alpha(3.7), -alpha(4.2), -(SEA)) and two non-deletions (alpha(Quong Sze), alpha(Constant Spring)). PCR pr...
journal_title：Journal of biotechnology
pub_type： 临床试验,杂志文章
doi：10.1016/j.jbiotec.2004.07.016
更新日期：2005-01-12 00:00:00
abstract：:European eel (Anguilla anguilla) elvers were intraperitoneally injected with different doses of 3,3',4,4'-tetrachlorobiyphenyl (PCB77) to examine and characterize the inductive effect of coplanar PCBs on CYP1A1 gene expression in liver and gills by using a semi-quantitative RT-PCR analysis. The influence of PCB77 inje...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2008.06.008
更新日期：2008-09-10 00:00:00
abstract：:Sulfite dissolving pulp from Eucalyptus grandis contained approximately 3.8% O-acetyl-4-O-methylglucuronoxylan with a molar ratio of xylose:4-O-methylglucuronic acid:acetyl group close to 13.6:1:6.2. The effects produced by purified endo-xylanases from two different glycosyl hydrolase families (family 10 and 11) as we...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(00)00324-2
更新日期：2000-10-13 00:00:00
abstract：:Proteolytically cleaved human 22 kDa growth hormone (22K hGH) between the amino acid residues 134 and 150 by plasmin or other proteases in vitro has been reported to be most active in growth promoting activity. In this study a deleted mutant hGH lacking amino acid residues from 135 to 146 and having more sensitivity t...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(99)00234-5
更新日期：2000-02-28 00:00:00
abstract：:Expanded bed adsorption chromatography is used to capture the protein product of interest from a crude biological suspension directly, thereby eliminating the need for the removal of the cell debris. While this technique may replace three or four unit operations in a typical downstream process for biological product r...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2007.07.724
更新日期：2008-02-01 00:00:00
abstract：:Transgenic hairy root (HR) systems constitute an interesting alternative to improve the efficiency of phytoremediation process. Since peroxidases (Px) have been associated with phenolic compounds removal, in the present work, transgenic tobacco HR, which expressed basic Px genes from tomato (tpx1 and tpx2), were estab...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2008.11.008
更新日期：2009-02-23 00:00:00
abstract：:Six bacterial strains with the capability of degrading textile dyes were isolated from sludge samples and mud lakes. Aeromonas hydrophila was selected and identified because it exhibited the greatest color removal from various dyes. Although A. hydrophila displayed good growth in aerobic or agitation culture (AGI cult...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(02)00303-6
更新日期：2003-02-27 00:00:00
abstract：:Signal peptides direct proteins to translocate across the bacterial cytoplasmic membrane. This study aimed to improve the level of extracellular secretion of recombinant carrageenase by recombining the gene encoding wild-type signal peptide (OmpZ) of Zobellia sp. ZM-2 κ-carrageenase into the expression vector pProEX-H...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.06.007
更新日期：2014-09-20 00:00:00
abstract：:The intrinsic kinetics of continuous yeast cell growth and ethanol production for a self-flocculating fusant yeast strain SPSC01 was investigated by means of mechanically dispersing the flocs and correspondingly established floc size distribution on-line monitoring technique using the focused beam reflectance measurem...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2005.12.029
更新日期：2006-07-13 00:00:00
abstract：:The genus Enterococcus occurs as native microbiota of fermented products due to its broad environmental distribution and its resistance to salt concentrations. Enterococcus faecalis F, a non-pathogenic strain isolated from a ripened cheese, has demonstrated useful enzymatic capabilities, a probiotic behavior and antib...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2018.06.346
更新日期：2018-10-10 00:00:00
abstract：:The fed-batch process for commercial production of riboflavin (vitamin B2) was optimized on-line using model-predictive control based on artificial neural networks (ANNs). The information required for process models was extracted from both historical data and heuristic rules. After each cultivation the process model w...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(00)00211-x
更新日期：2000-04-14 00:00:00
abstract：:This work explores some of the possibilities to replace human cytomegalovirus (CMV) core and/or enhancer promoter control elements to create new expression vectors for use with fish. The work is relevant to fish vaccination, since DNA vaccines use eukaryotic expression plasmids controlled by the human cytomegalovirus ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2012.08.005
更新日期：2012-12-15 00:00:00
abstract：:Gluconobacter oxydans can perform rapid incomplete oxidation of many sugars, sugar polyols and alcohols, and this outstanding ability shows a great potential in industrial bioconversion. Improvements of these industrially important strains would boost their productivities of important metabolites. However, the shortag...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.01.035
更新日期：2014-04-10 00:00:00
abstract：:We previously developed a novel one-step biotransformation process for the production of α-ketoglutarate (α-KG) from L-glutamic acid by a Bacillus subtilis whole-cell biocatalyst expressing an L-amino acid deaminase (pm1) of Proteus mirabilis. However, the biotransformation efficiency of this process was low owing to ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.07.431
更新日期：2014-10-10 00:00:00
abstract：:Efficient dynamic interactions among cofactor, enzymes and substrate molecules are of primary importance for multi-step enzymatic reactions with in situ cofactor regeneration. Here we showed for the first time that the above dynamic interactions could be significantly intensified by exerting an external alternating ma...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2013.05.016
更新日期：2013-10-20 00:00:00
abstract：:Chimeric virus-like particles (VLP) are known as promising tools in the development of safe and effective subunit vaccines. Recently, a technology platform to produce VLP based on the small surface protein (dS) of the duck hepatitis B virus was established. In this study, chimeric VLP were investigated displaying the ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2019.10.008
更新日期：2019-12-20 00:00:00
abstract：:The cyclic amino acid ectoine is a compatible solute serving as a protective substance against osmotic stress. Ectoine finds various applications due to its moisturizing effect. To avoid the disadvantages of the prevailing so-called "bacterial milking ectoine production process" caused by the high salt concentration, ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2017.04.039
更新日期：2017-09-20 00:00:00
abstract：:Biodegradable scaffolds are of great value in tissue engineering. We have developed a method for fabricating patient-specific vascular scaffolds from a biocompatible and biodegradable polymer, poly(L-lactide-co-epsilon-caprolactone). This method's usefulness is due to flexibility in the choice of materials and vascula...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2007.08.017
更新日期：2008-01-20 00:00:00
abstract：:A comparative study was made of Mung bean epoxide hydrolases-catalyzed asymmetric hydrolysis of styrene oxide to (R)-1-phenyl-1,2-ethanediol in an n-hexane/buffer biphasic system containing various hydrophilic ionic liquids (ILs). Compared to the n-hexane/buffer biphasic system alone, addition of a small amount of hyd...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2012.09.006
更新日期：2012-12-31 00:00:00
abstract：:Super-paramagnetic beads (SPMB)s used for a variety of molecular diagnostic assays are prepared by attaching pre-synthesized oligonucleotides to the surface via a cumbersome and low efficient method of carbodiimide-mediated amide bond formation. To mainstream the process, we describe a novel procedure of direct oligon...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2013.08.006
更新日期：2013-09-20 00:00:00
abstract：:The effect of an additional carbon source, lactate, on Penicillium camembertii growth on glutamate as both carbon and nitrogen sources was examined. Glutamate (and lactate) was present in excess in both media. Throughout the whole culture, similar growth time-courses were recorded on both media, indicating the absence...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2004.07.007
更新日期：2004-11-09 00:00:00
abstract：:Each of the twelve enzymes for glycolytic fermentation, eleven from Escherichia coli and one from Saccharomyces cerevisiae, have been over-expressed in E. coli and purified with His-tags. Simple assays have been developed for each enzyme and they have been assembled for fermentation of glucose to ethanol. Phosphorus-3...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2011.09.019
更新日期：2012-01-01 00:00:00
abstract：:The alpha-amylase (1, 4-alpha-d-glucanohydrolase; EC 3.2.1.1) and alpha-glucosidase (alpha-d-glucoside glucohydrolase; EC 3.2.1.20) secreted by Geobacillus thermodenitrificans HRO10 were purified to homogeneity (13.6-fold; 11.5% yield and 25.4-fold; 32.0% yield, respectively) through a series of steps. The molecular w...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2006.02.006
更新日期：2006-08-20 00:00:00
abstract：:To explore the utility of glycosyltransferases as novel biocatalysts, we isolated the glycosyltransferase genes CaUGT2 and SbUGTA1 from Catharanthus roseus and Starmerella bombicola, respectively and heterologously expressed them in Escherichia coli. The purified recombinant proteins were assayed with a variety of sma...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.10.011
更新日期：2015-12-20 00:00:00
abstract：:Enzyme prodrug therapies hold potential as a targeted treatment option for cancer patients. However, off-target effects can be detrimental to patient health and represent a safety concern. This concern can be alleviated by including a failsafe mechanism that can abort the therapy in healthy cells. This feature can be ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2017.09.005
更新日期：2017-10-20 00:00:00
abstract：:The combination of anaerobic and aerobic periods in the operation cycle of a Sequencing Batch Reactor (SBR) was chosen to study biological color removal from simulated textile effluents containing reactive, sulfonated, monoazo and diazo dyes, respectively, Remazol Brilliant Violet 5R and Remazol Black B. 90% color rem...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(01)00313-3
更新日期：2001-08-23 00:00:00
abstract：:The application of artificial chaperones in biotechnology has been inspired by the mechanism of molecular chaperones like GroEL/GroES. It involves addition of a capturing detergent during dilution of the chaotropic reagent, that prevents protein aggregation, and finally, addition of a oligosaccharide that removes the ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2009.01.023
更新日期：2009-03-25 00:00:00
abstract：:2'-Fucosyllactose (2-FL) is one of most abundant functional oligosaccharides in human milk, which is involved in many biological functions for human health. To date, most microbial systems for 2-FL production have been limited to use Escherichia coli JM strains since they cannot metabolize lactose. In this study, E. c...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.06.431
更新日期：2015-09-20 00:00:00
