Abstract:
:We previously showed that creation of a double strand DNA break (DSB) by expressing I-SceI in an engineered Trichoderma reesei (Hypocrea jecorina) strain containing a I-SceI recognition site improved transformation and homologous integration efficiencies. In this study, we further improved homologous integration frequencies by combining I-SceI mediated double strand break with disruption of the tku70 gene. The inability of the tku70 mutant to repair a I-SceI mediated DSB via NHEJ was used to force integration of an expression cassette with homologous flanks surrounding the DSB site. Besides expressing I-SceI from a plasmid, we also show that adding I-SceI enzyme during transformation was successful to generate DSBs. The I-SceI enzyme mediated integration, or SEMI, in combination with a Δtku70 mutant has a synergistic effect on homologous recombination efficiencies as 90-100% of the transformants exhibited integration of the expression cassette at the homologous site.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Ouedraogo JP,Arentshorst M,Nikolaev I,Barends S,Ram AFdoi
10.1016/j.jbiotec.2016.02.012subject
Has Abstractpub_date
2016-03-20 00:00:00pages
25-8eissn
0168-1656issn
1873-4863pii
S0168-1656(16)30057-8journal_volume
222pub_type
杂志文章abstract::New emerging technologies delivering benefits in terms of process robustness and economy are an inevitable prerequisite for monoclonal antibody purification processes intensification. Caprylic acid was proven as an effective precipitating agent enabling efficient precipitaton of product- and process-related impurities...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2018.05.001
更新日期:2018-08-10 00:00:00
abstract::A PHA producing microbial culture dominated by Plasticicumulans acidivorans was enriched in a pilot plant using fermented wastewater from the Mars candy bar factory. The pilot plant comprised (1) anaerobic fermentation, (2) enrichment of a PHA-producing microbial community and (3) accumulation for maximization of the ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2014.10.022
更新日期:2014-12-20 00:00:00
abstract::Pimprinine, an extracellular alkaloid has been isolated from the culture filtrate of Streptomyces CDRIL-312. Pimprinine was subsequently purified using silica gel column chromatography and also by preparatory thin layer chromatography. Some physicochemical properties, antimicrobial activities (in-vitro) and pharmacolo...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/s0168-1656(01)00244-9
更新日期:2001-06-01 00:00:00
abstract::A gene encoding for hirudin, a potent thrombin inhibitor, was expressed in Escherichia coli, which is the most widely used host. When the recombinant hirudin analog, CX-397, was overproduced by E. coli (600 mg l(-1)) in the absence of nutrient amino acids in the culture medium, the presence of two derivatives in the f...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/s0168-1656(01)00396-0
更新日期:2002-02-14 00:00:00
abstract::Enzymatic synthesis of acylated derivatives of a monosaccharidic flavonoid chrysoeriol-7-O-beta-D-(3''-E-p-coumaroyl)-glucopyranoside as well as of a disaccharidic flavonoid chrysoeriol-7-[6'''-O-acetyl-beta-D-allosyl-(1-->2)-beta-D-glucopyranoside], isolated from Greek endemic plants, was performed using an immobiliz...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2004.12.002
更新日期:2005-03-30 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
abstract::To improve tumor-to-tissue ratios of anticancer agents in radioimmunotherapy, a three-step targeting approach was used to deliver biotinylated liposomes to human ovarian cancer cells (NIH:OVCAR-3, SK-OV-3) in vitro. Targeting was based upon the use of two antibodies specific for the CA-125 antigen that is highly expre...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/s0168-1656(01)00424-2
更新日期:2002-03-28 00:00:00
abstract::A fibrous-bed bioreactor containing the coculture of Pseudomonas putida and P. fluorescens immobilized in a fibrous matrix was developed to degrade benzene (B), toluene (T), ethylbenzene (E), and o-xylene (X) in synthetic waste streams. The kinetics of BTEX biodegradation by immobilized cells adapted in the fibrous-be...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/s0168-1656(98)00166-7
更新日期:1999-01-22 00:00:00
abstract::Proteins are multifunctional: their amino acid sequences simultaneously determine folding, function and turnover. Correspondingly, evolution selected for compromises between rigidity (stability) and flexibility (folding/function/degradation), to the result that generally the free energy of stabilization of globular pr...
journal_title:Journal of biotechnology
pub_type: 杂志文章,评审
doi:10.1016/s0168-1656(00)00236-4
更新日期:2000-05-26 00:00:00
abstract::Norovirus infections cause severe gastroenteritis in millions of people every year. Infection requires the recognition of histo-blood group antigens (HBGAs), but such interactions can be inhibited by human milk oligosaccharides (HMOs), which act as structurally-similar decoys. HMO supplements could help to prevent nor...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2020.05.001
更新日期:2020-07-20 00:00:00
abstract::We established a novel method to analyze the gene expression of prokaryotes by modifying and optimizing the HiCEP (high coverage gene expression analysis) method, which was originally developed for eukaryotic gene expression profiling. Following the removal of abundant rRNA, the mRNA of prokaryotic cells was enzymatic...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2005.11.004
更新日期:2006-05-29 00:00:00
abstract::Kinetic models have a great potential for metabolic engineering applications. They can be used for testing which genetic and regulatory modifications can increase the production of metabolites of interest, while simultaneously monitoring other key functions of the host organism. This work presents a methodology for in...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2016.01.005
更新日期:2016-03-20 00:00:00
abstract::A transgenic tobacco plant (Nicotiana tabacum L.) expressing a fungal pectin methylesterase (PME; EC 3.1.1.11) gene derived from a black filamentous fungus, Aspergillus niger was created. Fungal PME should have a wider range of adaptability to substrate pectin compared with plant PME. As expected, the proportion of me...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2004.04.015
更新日期:2004-08-05 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 gene encoding histamine dehydrogenase in Rhizobium sp. 4--9 has been cloned and overexpressed in Escherichia coli. The coding region of the gene was 2,079 bp and encoded a protein of 693 amino acids with a calculated molecular mass of 76,732 Da. This histamine dehydrogenase was related to histamine dehydrogenase f...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2005.04.005
更新日期:2005-09-29 00:00:00
abstract::Poly-l-cysteine (PLCys) is drawing attention as a potential sorbent of thiol (SH)-reactive toxic heavy metal ions in the wastewater and polluted soils. However, preparation of PLCys relies on chemically synthesized polymers, in which SH groups must be protected and deprotected prior to use. On the other hand, α-chymot...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2011.12.021
更新日期:2012-02-10 00:00:00
abstract::Yarrowia lipolytica can accumulate large amounts of storage lipids and has considerable potential for the production of polyunsaturated fatty acids and other lipids for biofuels. When the nitrogen source is exhausted in the medium, the key intermediate, citrate, is converted to acetyl-CoA by ATP:citrate lyase (ACL) fo...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2014.10.004
更新日期:2014-12-20 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::Microbiologically induced calcium carbonate precipitation (MICCP) is a naturally occurring biological process that has shown its potential in remediation of a wide range of structural damages including concrete cracks. In this study, genetically engineered microorganisms, capable of producing extracellular polymeric s...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2012.07.001
更新日期:2012-10-31 00:00:00
abstract::Generation and characterization of knockout clones is a widely used approach to evaluate the specific function of a gene product in Dictyostelium discoideum. The mutant clones are generally obtained by double homologous recombination in the target gene. A frequent limitation to obtaining mutants is the low frequency o...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2005.08.023
更新日期:2006-03-09 00:00:00
abstract::In this report, we describe the results of a rat pituitary cell culture experiment done on STS-65 in which the effect of cell feeding on the release of the six anterior pituitary hormones was studied. We found complex microgravity-related interactions between the frequency of cell feeding and the quantity and quality ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/0168-1656(96)01373-9
更新日期:1996-06-27 00:00:00
abstract::Owing to its dual impact on tissue engineering (neovascularization of tissue implants) and cancer treatment (prevention of tumor-induced vascularization), management and elucidation of vascularization phenomena remain clinical priorities. Using a variety of primary human cells and (neoplastic) cell lines assembled in ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2005.03.016
更新日期:2005-08-04 00:00:00
abstract::Cannabis sativa L. plants produce a diverse array of secondary metabolites. Cannabis cell cultures were treated with biotic and abiotic elicitors to evaluate their effect on secondary metabolism. Metabolic profiles analysed by (1)H NMR spectroscopy and principal component analysis (PCA) showed variations in some of th...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2009.05.006
更新日期:2009-08-20 00:00:00
abstract::Delivery of macromolecules into the cytosolic space of eukaryotic cells is a pressing challenge in biopharmaceutics. Macromolecules are often encapsulated into liposomes for protection and improved distribution, but the their size often induces endocytosis of the vehicle at the target site, leading to degradation of t...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2013.01.029
更新日期:2013-04-15 00:00:00
abstract::The kinetics of microbial growth and the biodegradation of methanol and toluene in (a) biofilters (BFs), and (b) biotrickling filters (BTFs), packed with inert materials, has been studied and analyzed. The specific growth rate, mu, for the treatment of methanol was 0.037h(-1) for a wide range of operating conditions. ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2008.08.001
更新日期:2008-11-25 00:00:00
abstract::The extraction of 13-hydroperoxide-lyase activity from mint leaves as well as its use for C6-aldehyde production was studied in this work. The enzyme cleaves 13(S)-hydroperoxy-C18 fatty acids into C6-aldehyde and C12-oxo-acid. Two mint species were tested: Mentha veridis and Mentha pulegium. The headspace injection me...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2004.03.005
更新日期:2004-07-01 00:00:00
abstract::Reteplase is the third generation of thrombolytic medicine and has many advantages over commercial t-PA. However, over-expressing recombinant reteplase in E. coli always accumulates as inclusion bodies due to nine pairs of disulfide bonds formation that is the main obstacle for correct folding. In this paper, in order...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2014.10.028
更新日期:2014-12-20 00:00:00
abstract::We constructed a plasmid, designated pNPP126, containing a DNA sequence encoding a fusion protein composed of Bacillus amyloliquefaciens neutral protease prepeptide (signal peptide) and human pancreatic secretory trypsin inhibitor (hPSTI), where the mature hPSTI is accurately fused to the 3'-terminal of the prepeptide...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/0168-1656(92)90094-p
更新日期:1992-04-01 00:00:00
abstract::Procedures were developed for 'self-immobilisation' of Solanum aviculare cells to eliminate the need for artificial immobilisation supports. Depending on the cytokinin used in liquid medium, compact aggregates 0.4-2.0 cm in diameter were formed without dispersed cells also being present. Histochemical analysis showed ...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/0168-1656(91)90077-9
更新日期:1991-06-01 00:00:00
abstract::Three of the most abundant proteins (OmpW, MopB and SodM) of the extracellular proteome of Xanthomonas campestris pv. campestris were analysed in a luminol-based oxidative burst assay to identify novel pathogen-associated molecular patterns (PAMP). Tobacco cell suspension cultures were used as a model system to monito...
journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2006.02.022
更新日期:2006-10-20 00:00:00