Abstract:
:The development of SIN-lentivectors has paved the way for the double copy vectors (DCV), which substitute the deletion in the 3'LTR with either transgenic or insulator sequences. However, the limits of this approach remain unclear. Previous results have demonstrated that transduction efficiencies of DCV carrying large insulator inserts in their 3'LTRs were impaired in a size-dependent manner. We wondered if this was also true for promoter-transgene inserts and whether they remained functional upon integration into the genome. Therefore, we designed a series of DCV with increasing 3'LTR sizes containing different promoter-transgene combinations. Transduced cells were scored for both transduction efficiency and insert functionality. We found that the transduction efficiencies indeed were impaired in a size-dependent way. Efficiency with inserts below 1 kb linearly decreased with size, while sizes between 1 and 2 kb showed a further decrease to a minimum of 5% of an original "empty" SIN-vector. However, we did not find an LTR size that completely abolished transduction. Moreover, we demonstrated that all inserts remained functional regardless the promoter-transgene combination used. Therefore, we conclude from our data that DCV indeed remain functional, but transduction efficiencies drop radically when inserts larger than 1 kb are being used.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Bos TJ,De Bruyne E,Van Lint S,Heirman C,Vanderkerken Kdoi
10.1016/j.jbiotec.2010.07.010subject
Has Abstractpub_date
2010-10-01 00:00:00pages
37-40issue
1eissn
0168-1656issn
1873-4863pii
S0168-1656(10)00328-7journal_volume
150pub_type
杂志文章abstract::Lactulose and galacto-oligosaccharides (GOS) are well recognized prebiotics derived from lactose. In the synthesis of lactulose with β-galactosidases GOS are also produced, but the ratio of lactulose and GOS in the product can be tuned at will, depending on the operation conditions, so to obtain an optimal product dis...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2015.06.394
更新日期:2015-09-10 00:00:00
abstract::Stem borer resistant transgenic parental lines, involved in hybrid rice, were produced by Agrobacterium-mediated gene transfer method. Two pSB111 super-binary vectors containing modified cry1Ab/cry1Ac genes driven by maize ubiquitin promoter, and herbicide resistance gene bar driven by cauliflower mosaic virus 35S pro...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2004.04.004
更新日期:2004-07-15 00:00:00
abstract::The partitioning of proteins and other biomaterials between two aqueous phases containing polyethyleneglycol and dextran is a strong function of the molecular weight of the two polymers. Although both polymers are polydispersed (especially Dx) most theoretical treatments refer only to the average molecular weight (num...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/0168-1656(91)90224-j
更新日期:1991-09-01 00:00:00
abstract::Aliphatic thiols are effective as redox buffers for folding non-native disulfide-containing proteins into their native state at high pH values (8.0-8.5) but not at neutral pH values (6-7.5). In developing more efficient and flexible redox buffers, a series of aromatic thiols was analyzed for its ability to fold scramb...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2004.09.005
更新日期:2005-02-09 00:00:00
abstract::Corynebacterium glutamicum produces succinate from glucose via the reductive tricarboxylic acid cycle under microaerobic and anaerobic conditions. We identified a NCgl2130 gene of C. glutamicum as a novel succinate exporter that functions in succinate production, and designated sucE1. sucE1 expression levels were high...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2011.03.010
更新日期:2011-06-10 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::Porcine muscle adenylate kinase (ADK) was overproduced in Escherichia coli using the expression plasmid with double A-T-G codon at the translational starting site and the Shine-Dalgarno (SD) sequence 10 bp apart from the first A-T-G. We used the expression vectors pKK223-3 and pMK2. pMK2 is about 10-20 times larger in...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/0168-1656(94)90176-7
更新日期:1994-02-14 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::Combining the isochorismate synthase EntC and the chorismatase FkbO in a sequential enzyme cascade provides a useful system for the biocatalytic production and subsequent purification of isochorismate from an isochorismate/chorismate mixture. FkbO has a strict preference for chorismate - isochorismate is not accepted ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2014.06.003
更新日期:2014-12-10 00:00:00
abstract::Enterokinase, a two-chain duodenal serine protease, activates trypsinogen by removing its N-terminal propeptide. Due to a clean cut after the non-primed site recognition sequence, the enterokinase light chain is frequently employed in biotechnology to separate N-terminal affinity tags from target proteins with authent...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2013.10.022
更新日期:2013-12-01 00:00:00
abstract::Recently, some research in the field of enhanced biological phosphorus removal (EBPR) has been focused on studying systems where the electron donor (substrate) and the electron acceptor (nitrate or oxygen) are present simultaneously. This can occur, for example, in a full scale wastewater treatment plant during heavy ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2005.10.018
更新日期:2006-05-03 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::A continuous enzymatic process for the production of the prebiotic disaccharide lactulose through transgalactosylation was developed using free and immobilized beta-glycosidase from Pyrococcus furiosus. The hyperthermostable beta-glycosidase (CelB) was immobilized onto an anion-exchange resin (Amberlite IRA-93) or ont...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2009.12.017
更新日期:2010-02-15 00:00:00
abstract::4-vinylguaiacol, a kind of volatile phenolic compound with tobacco flavor, is widely used as a component of edible flavor and intermediate of medicine. Ferulic acid is usually used as substrate for the biosynthesis of 4-vinylguaiacol. However, the price of ferulic acid is high, leading to high production cost. In this...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2018.07.021
更新日期:2018-09-10 00:00:00
abstract::The PRNP gene encodes a cellular protein named prion, whose misfolded form has been implicated in a number of neuropathic diseases in mammals such as the Bovine Spongiform Encephalopathy (BSE) in cattle. BSE has brought devastating impact on the world economy and human health. Recently, several groups have performed t...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2012.10.018
更新日期:2013-02-20 00:00:00
abstract::Epigallocatechin-3-O-gallate (EGCg) and related polyphenolic compounds found in tea are known to have antioxidative activities. However, they also have pro-oxidative activities such as generation of hydrogen peroxide. In this report, we investigated the effect on cells and showed the potential usage of EGCg in cell pr...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2006.08.006
更新日期:2007-01-20 00:00:00
abstract::This study examined the kinetics and substrate selectivity of a GH11 Bacillus subtilis XynA xylanase (BsX) sensitive to inhibition by TAXI and an engineered variant, which is much less inhibited by TAXI (BsX(mut)). The main purpose of the work was to elucidate any influence of the structural point mutations on the kin...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2010.02.012
更新日期:2010-04-15 00:00:00
abstract::Rhizomucor miehei lipase (RML), a GRAS catalyst with wide applications, was overexpressed in Yarrowia lipolytica, also a GRAS unconventional yeast, via a combined strategy, optimization for promoter, gene dosage and fermentation process. The lipase activity of the recombinant strain was first increased from 19.5 to 26...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2019.09.004
更新日期:2019-12-20 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::Pituitary tumor-transforming gene 1 (PTTG1), a proto-oncogene, is associated with tumor formation, proliferation and invasiveness. F-PBF(beta-TrCP), a fusion protein, was produced by replacing the WD40-repeat of F-box protein beta-TrCP with the PTTG1-binding factor (PBF) for targeted degradation of PTTG1. To evaluate ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2008.09.004
更新日期:2009-01-01 00:00:00
abstract::The grapevine genetic transformation programs have relayed on the use of solid media-based somatic embryogenesis. To reach a high throughput of candidate gene evaluation in 'Thompson Seedless', a semi-automatic system allowing viable transformation of explants was designed. An intermediate procedure using liquid media...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2008.09.009
更新日期:2009-01-01 00:00:00
abstract::This review is aimed at biochemists and molecular biologists, and covers the chemistry and key features involved in the solid-phase synthesis of a variety of the better known DNA and RNA analogues by the phosphoramidite and H-phosphonate methods. A wide spectrum of biological applications such as inhibition of gene ex...
journal_title:Journal of biotechnology
pub_type: 杂志文章,评审
doi:10.1016/0168-1656(95)00012-f
更新日期:1995-07-31 00:00:00
abstract::The elemental composition of microorganisms varies widely with respect to the N, S, P and ash content as well as many trace elements depending on microorganism and growth conditions. As a consequence, the mass of 1 C-mol of biomass can differ considerably from one microorganism to another. Experimental results show th...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/0168-1656(95)00135-0
更新日期:1995-12-01 00:00:00
abstract::In the two-step fermentative production of vitamin C, its precursor 2-keto-L-gulonic acid (2-KLG) was synthesized by Ketogulonicigenium vulgare through co-culture with Bacillus megaterium. The reconstruction of the amino acid metabolic pathway through completed genome sequence annotation demonstrated that K. vulgare w...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2011.08.007
更新日期:2011-12-10 00:00:00
abstract::Gene farming is one of the most promising areas in modern biotechnology. To assay the potential usefulness of transgenic rabbits as bioreactors, one call embryos were microinjected with a chimeric gene comprising 5' sequences from mouse whey acidic protein gene (mWAP) linked to the human growth hormone (hGH) gene. Tra...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/0168-1656(95)00026-m
更新日期:1995-05-15 00:00:00
abstract::The complete genome sequence of Streptomyces fulvissimus (DSM 40593), consisting of a linear chromosome with a size of 7.9Mbp, is reported. Preliminary data indicates that the chromosome of S. fulvissimus contains 32 putative gene clusters involved in the biosynthesis of secondary metabolites, two of them showing very...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2013.08.013
更新日期:2013-10-10 00:00:00
abstract::Paramagnetic carriers, which are linked to antibodies enable highly specific biological cell separations. With the colloidal synthesis of superparamagnetic Co and FeCo nanocrystals with superior magnetic moments the question about their potential to replace magnetite as the magnetically responsive component of magneti...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2004.04.019
更新日期:2004-08-26 00:00:00
abstract::Lentinula edodes, the popular shiitake mushroom, is one of the most important cultivated edible mushrooms. It is used as a food and for medicinal purposes. Here, we present the 46.1 Mb draft genome of L. edodes, comprising 13,028 predicted gene models. The genome assembly consists of 31 scaffolds. Gene annotation prov...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2016.02.032
更新日期:2016-04-10 00:00:00
abstract::Escherichia coli overexpressing a NAD(+)-dependent formate dehydrogenase (FDH) from Candida boidinii was grown in chemostat culture on various carbon sources at 0.05 h(-1) dilution rate, under anaerobic conditions using defined medium and compared to a control without the heterologous FDH pathway. Metabolic fluxes, NA...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2005.02.006
更新日期:2005-06-29 00:00:00
abstract::A new and simple method was devised to obtain immunoglobulin VH genes directly from primary cultured B cells specific for a short peptide. Peptide-specific B cells were separated from splenocytes of peptide-immunized BALB/c mice with antigen-coated magnetic beads, and were cloned by a limitedly diluted culture in the ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/0168-1656(96)01506-4
更新日期:1996-08-20 00:00:00