Abstract：
:The objective of this study was to identify and isolate endogenous promoters in Chinese hamster ovary (CHO) cells using a promoter trap approach. A promoter-less vector harboring a green fluorescent protein (GFP)-hygromycin resistance gene cassette was designed and transfected into CHO cells. Putative promoters were identified by selecting for GFP(+) clones under hygromycin selection. Genomic DNA from these clones was then digested and self-ligated to give rise to a plasmid carrying the putative promoter sequence as well as elements for replication in E. coli. Functional promoter sequences were subsequently identified by screening the recovered plasmids for their ability to drive GFP expression upon re-transfection into CHO cells. One of the fragments isolated through this approach was found to drive gene expression in two different reporter systems. Further dissection of the fragment led to the identification of a 156-bp element that was four-fold more active than the full-length fragment and 66% as active as the SV-40 promoter. Thus, promoter trap represents an effective strategy for identifying endogenous regulatory regions that can potentially be incorporated into expression vectors to augment expression of recombinant biopharmaceuticals.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Chen J,Haverty J,Deng L,Li G,Qiu P,Liu Z,Shi Sdoi
10.1016/j.jbiotec.2013.07.001subject
Has Abstractpub_date
2013-09-10 00:00:00pages
255-61issue
3eissn
0168-1656issn
1873-4863pii
S0168-1656(13)00281-2journal_volume
167pub_type
杂志文章abstract：:Two cyanobacterial strains, Synechocystis sp. PCC 6803 and Nostoc punctiforme ATCC 29133 were immobilized within magnesium phosphate based cements, showing a viability and activity for at least 4 weeks. These biohybrids are considered as an alternative photobioreactor material for bioremediation or an improved yield o...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2016.02.024
更新日期：2016-04-10 00:00:00
abstract：:AL (SpA A domain-PpL B3 domain), LD5 (PpL B3 domain-SpA D domain-PpL B3 domain-SpA D domain-PpL B3 domain, L-D-L-D-L) and LD3 (PpL B3 domain-SpA D domain-PpL B3 domain, L-D-L) are novel evolved Ig binding molecules (NEIBMs) derived from the in vitro molecular evolution of combinatorial phage libraries displaying rando...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.05.014
更新日期：2014-08-20 00:00:00
abstract：:Thioalkalivibrio versutus D301 isolated from Soda Lake is a haloalkaliphilic and obligated chemolithoautotrophic Gram-negative bacterium. The strain has a good adaption to hyperhaline and highly alkaline environment and a powerful sulfur-oxidizing ability. Here, we present the complete genome sequence of T. versutus D...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2016.04.019
更新日期：2016-06-10 00:00:00
abstract：:Methods of adapting micro-organisms to an inhibiting factor in an active industrial bioprocess were examined with an acetic acid fermentation as model. With the aim of automatic control, a fuzzy-logic system was developed on the basis of the collected knowledge of skilled vinegar brewers. In a first step, this fuzzy s...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(02)00065-2
更新日期：2002-08-07 00:00:00
abstract：:The efficiency of protein synthesis for glyceraldehyde-3-phosphate dehydrogenase (G3PDH) was examined with several in vitro coupled transcription/translation protein synthesis systems based on Escherichia coli lysate, wheat germ, or reticulocyte lysate, and an in vitro translation system based on wheat germ extract. A...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2007.08.008
更新日期：2008-01-20 00:00:00
abstract：:When subjected to a Picea abies suspension cell culture, beta-pinene, either one of the pure enantiomers or the racemate, was transformed mainly to trans-pinocarveol along with the minor products myrtenol, alpha-terpineol, pinocarvone, myrtenal and cis-pinocarveol. The absolute configuration of the major products corr...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2003.10.009
更新日期：2004-01-22 00:00:00
abstract：:Oxygen is a key substrate in animal cell metabolism. It has been reported that the oxygen uptake rate (OUR) is a good indicator of cellular activity, and even under some conditions, a good indicator of the number of viable cells. The measurement of OUR is difficult due to many different reasons. In particular, the ver...
journal_title：Journal of biotechnology
pub_type： 杂志文章,评审
doi：10.1016/s0168-1656(98)00046-7
更新日期：1998-08-12 00:00:00
abstract：:A hydrogen electrode based on a hydrogenase immobilized on a carbon material has been developed. The equilibrium hydrogen potential was reached in an atmosphere of H2 on the enzyme electrode according to the mechanism of direct exchange of electrons between the electrode and the enzyme active center. The electrochemic...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(93)90095-5
更新日期：1993-02-01 00:00:00
abstract：:Peptide synthesis was carried out in a variety of organic solvents with low contents of water. The enzyme was deposited on the support material, celite, from an aqueous buffer solution. After evaporation of the water the biocatalyst was suspended in the reaction mixtures. The chymotrypsin-catalyzed reaction between Z-...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(90)90090-x
更新日期：1990-09-01 00:00:00
abstract：:Glucose oxidase (GOD) was immobilized on cellulose acetate-polymethylmethacrylate (CA-PMMA) membrane. The immobilized GOD showed better performance as compared to the free enzyme in terms of thermal stability retaining 46% of the original activity at 70 degrees C where the original activity corresponded to that obtain...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2005.08.019
更新日期：2006-02-10 00:00:00
abstract：:Porcine circovirus Cap protein production by P. pastoris with strong AOX promoter suffered with the problems with traditional pure methanol induction: (1) inefficient methanol metabolism; (2) extensive oxygen supply load; (3) difficulty in stable DO control; (4) low protein titer. In this study, based on the differenc...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.02.018
更新日期：2014-05-10 00:00:00
abstract：:The aim of this study was to improve the ability of Aspergillus terreus lipase to separate the racemic ketoprofen vinyl ester into individual enantiomers using hollow self-assembly alginate-graft-poly(ethylene glycol)/α-cyclodextrins (Alg-g-PEG/α-CD) spheres as enzyme immobilization carriers. The morphology and size o...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.11.032
更新日期：2015-01-20 00:00:00
abstract：:beta-galactosidase reporter plasmids containing different viral or minimal promoters are commonly used to correct variable transfection efficiencies in transient transfection experiments. The transcriptional activity of these promoters is thought to be stable under most circumstances. To determine if expression of bet...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(01)00277-2
更新日期：2001-07-12 00:00:00
abstract：:Many recent epigenetic studies utilize the advantages of CRISPR/dCas9 based tools in linking certain epigenetic modification with gene expression regulation. Various multifactorial diseases often contain changed epigenetic signatures at many loci, so tools for simultaneously targeting different loci would significantl...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2019.05.306
更新日期：2019-08-10 00:00:00
abstract：:The adsorption behaviors of two glucanases, TvEG and TrCel7A, on three lignins were investigated. Three lignins were isolated from raw aspen and its pretreated solid residue. The isolated lignins were labeled as Asp-MWL, DA-MWL (pretreated by dilute acid), and GL-MWL (pretreated by green liquor), respectively. The sur...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2020.07.013
更新日期：2020-11-10 00:00:00
abstract：:Linoleate 13-hydratase from Lactobacillus acidophilus LMG 11470 converted linoleic acid to hydroxyl fatty acid, which was identified as 13S-hydroxy-9(Z)-octadecenoic acid (13-HOD) by GC-MS and NMR. The expression of linoleate 13-hydratase gene in Escherichia coli was maximized by using pACYC plasmid and super optimal ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.05.006
更新日期：2015-08-20 00:00:00
abstract：:The enzymatic bioconversion of chitin is of considerable interest for the natural production of bioactive compounds such as chitooligosaccharides and N-acetyl-d-glucosamine (GlcNAc). Key enzymes are involved in the natural processing of chitin, hydrolyzing this abundant biopolymer to yield chitooligosaccharides with s...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.12.038
更新日期：2016-02-20 00:00:00
abstract：:Deracemization of D,L-phosphinothricin (D,L-PPT) is one of the most promising routes for preparation of optically pure L-PPT. In this work, an efficient multi-enzyme redox cascade was developed for deracemization ofPPT, which includes oxidative reaction and reductive reaction. The oxidative reaction catalyzing oxidati...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2020.09.024
更新日期：2021-01-10 00:00:00
abstract：:In the current study, a three-tiered mutagenesis strategy was employed to simultaneously improve the thermostability and activity of halohydrin dehalogenase from Agrobacterium radiobacter AD1 (HheC) by engineering the last ten amino acids (Met245∼Glu254) of its C-terminal region. Initially, truncated mutagenesis resul...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.08.013
更新日期：2015-10-20 00:00:00
abstract：:A novel bacterial species was isolated from an industrial-scale biogas plant. The isolate Herbinix hemicellulosilytica T3/55(T) is able to degrade crystalline cellulose. Comparative 16S rRNA gene sequencing demonstrated that the isolate is closely related to environmental samples forming a hitherto unknown sub-cluster...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.07.022
更新日期：2015-11-20 00:00:00
abstract：:The synthesis of methyl alpha-D-glucooligosaccharides, using sucrose as glucosyl donor and methyl alpha-D-glucopyranoside as acceptor, was studied with dextransucrase from Leuconostoc mesenteroides NRRL B-1299. The enzyme was immobilized by entrapment in alginate. By NMR and mass spectrometry we identified three homol...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2005.12.031
更新日期：2006-07-13 00:00:00
abstract：:Baculovirus vectors show promise as a novel tool for gene delivery into mammalian cells and gene transfer with wild-type baculovirus has been demonstrated both in vitro and in vivo. To study expression and intracellular trafficking of foreign viral membrane proteins in baculovirus-transduced mammalian cells, the envel...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2004.06.015
更新日期：2004-10-19 00:00:00
abstract：:In the present study we show that recombinant bacterial CotA-laccase from Bacillus subtilis is able to decolourise, at alkaline pH and in the absence of redox mediators, a variety of structurally different synthetic dyes. The enzymatic biotransformation of the azo dye Sudan Orange G (SOG) was addressed in more detail ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2008.09.001
更新日期：2009-01-01 00:00:00
abstract：:Cancer immunotherapy is a promising intervention to fight against this global health problem. In particular targeting immune checkpoints, such as cytotoxic T-lymphocyte associated antigen-4 (CTLA-4) and programmed-death protein 1 (PD-1), by specific monoclonal antibodies is a current treatment for many malignances. A ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2021.01.016
更新日期：2021-01-21 00:00:00
abstract：:Porcine circovirus type 2 (PCV2) is the primary causative agent of an economically important swine disease, now known as porcine-associated disease (PCVAD). The only structural protein of viral capsid, Cap has become the major target for development of PCV2 subunit vaccines. The purpose of this study is to express Cap...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.04.006
更新日期：2014-07-10 00:00:00
abstract：:Oleaginous yeast Trichosporon cutaneum demonstrated excellent lipid accumulation performance and inhibitor tolerance derived from lignocellulose pretreatment. Here we firstly report a 30.45Mb assembly genome of T. cutaneum ACCC 20271 for understanding the microbial lipid biosynthesis and mechanism of inhibitor toleran...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2016.04.043
更新日期：2016-06-20 00:00:00
abstract：:Functional screening for lipolytic enzymes from a metagenomic library (origin: Jae Sawn hot spring, Thailand) resulted in isolation of a novel patatin-like phospholipase (PLP) and an esterase (Est1). PLP contained four conserved domains similar to other patatin-like proteins with lipid acyl hydrolase activity. Likewis...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2007.08.046
更新日期：2008-01-01 00:00:00
abstract：:Acetogenic bacteria like Acetobacterium woodii represent an ancient group of anaerobic microorganisms which use hydrogen and carbon dioxide to produce acetate. Cell concentrations and space-time yields are usually low in gas fermentations. A standard stirred‑tank bioreactor with continuous gas supply was equipped with...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.07.020
更新日期：2015-10-20 00:00:00
abstract：:A serum-dependent and two serum-independent variants of the Bowes melanoma cell line, RPMI7272, were transfected with plasmids containing a geneticin-resistance (neo) gene transcribed by the HSV thymidine kinase promoter and an SV40 T antigen gene under control of the mouse metallothionein I promoter. T-antigen increa...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(92)90014-z
更新日期：1992-11-01 00:00:00
abstract：:Enzyme engineering by molecular modelling and site-directed mutagenesis can be remarkably efficient. Directed enzyme evolution appears as a more general strategy for the isolation of catalysts as it can be applied to most chemical reactions in aqueous solutions. Selections, as opposed to screening, allow the simultane...
journal_title：Journal of biotechnology
pub_type： 杂志文章,评审
doi：10.1016/j.jbiotec.2004.03.032
更新日期：2004-09-30 00:00:00
