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Registros recuperados: 12
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A biolistic process for in vitro gene transfer into chicken embryos BJMBR
Ribeiro,L.A.; Mariani,P.D.S.C.; Azevedo,J.L.; Rech,E.L.; Schmidt,G.S.; Coutinho,L.L..
Chicken embryos kept in culture medium were bombarded using a high helium gas pressure biolistic device. To optimize the factors that affect transformation efficiency, the lacZ gene under control of the human cytomegalovirus immediate early enhancer/promoter was used as a reporter gene. There was an inverse relationship between survival rate and transformation efficiency. The best conditions obtained for high embryo survival and high transformation efficiency were achieved with 800 psi helium gas pressure, 500 mmHg vacuum, gold particles, an 8 cm DNA-coated microparticle flying distance to the embryo and embryo placement 0.5 cm from the center of the particle dispersion cone. Under these conditions, transformation efficiency was 100%, survival rate 25% and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Biolistic process; Chicken embryo; Gene transfer; SS-galactosidase; Green fluorescent protein; GFP.
Ano: 2001 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2001000900003
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Asparaginase II-GFP fusion as a tool for studying the secretion of the enzyme under nitrogen starvation BJM
Sotero-Martins,Adriana; Bon,Elba Pinto da Silva; Carvajal,Elvira.
Production of asparaginase II of Saccharomyces cerevisiae is regulated by nitrogen and can be used as a model system for studying other secreted proteins in yeast. Green fluorescent protein (GFP) from Aequorea victoria was fused to the carboxy-terminus of the enzyme by genomic integration to the locus ASP3 of S. cerevisiae. We determined asparaginase II activity, mRNA ASP3, mRNA ASP3-GFP and GFP fluorescence. Nitrogen starvation in cells carrying the chimera ASP3-GFP caused an increase in fluorescence and in the expression of ASP3. We have shown that cells producing the chimera Asp3-GFPp displayed the same response to nitrogen starvation as control cells. We demonstrated that Asp3-GFPp can be used for studying asparaginase II secretion under nitrogen...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Asparaginase II; Nitrogen regulation; Saccharomyces cerevisiae; Genomic integration; Green fluorescent protein.
Ano: 2003 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822003000400017
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Characterization of a small cryptic plasmid from endophytic Pantoea agglomerans and its use in the construction of an expression vector Genet. Mol. Biol.
Rudi Emerson de Lima,Procópio; Araújo,Welington Luiz; Andreote,Fernando Dini; Azevedo,João Lúcio.
A circular cryptic plasmid named pPAGA (2,734 bp) was isolated from Pantoea agglomerans strain EGE6 (an endophytic bacterial isolate from eucalyptus). Sequence analysis revealed that the plasmid has a G+C content of 51% and contains four potential ORFs, 238(A), 250(B), 131(C), and 129(D) amino acids in length without homology to known proteins. The shuttle vector pLGM1 was constructed by combining the pPAGA plasmid with pGFPmut3.0 (which harbors a gene encoding green fluorescent protein, GFP), and the resulting construct was used to over-express GFP in E. coli and P. agglomerans cells. GFP production was used to monitor the colonization of strain EGE6gfp in various plant tissues by fluorescence microscopy. Analysis of EGE6gfp colonization showed that 14...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Green fluorescent protein; Bacteria; Host plant; Eucalyptus.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000100018
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First evidence for a Vibrio strain pathogenic to Mytilus edulis altering hemocyte immune capacities ArchiMer
Ben Cheikh, Yosra; Travers, Marie-agnes; Morga, Benjamin; Godfrin, Yoann; Rioult, Damien; Le Foll, Franck.
Bacterial isolates were obtained from mortality events affecting Mytilus edulis and reported by professionals in 2010-2013 or from mussel microflora. Experimental infections allowed the selection of two isolates affiliated to V. splendidus/V. hemicentroti type strains: a virulent 10/068 1T1 (76,6% and 90% mortalities in 24h and 96h) and an innocuous 12/056 M24T1 (0% and 23,3% in 24h and 96h). These two strains were GFP-tagged and validated for their growth characteristics and virulence as genuine models for exposure. Then, host cellular immune responses to the microbial invaders were assessed. In the presence of the virulent strain, hemocyte motility was instantaneously enhanced but markedly slowed down after 2h exposure. By contrast, hemocyte velocity...
Tipo: Text Palavras-chave: Innate immunity; Molluscs; Cell-mediated immune response; Bacterial infections; Green fluorescent protein.
Ano: 2016 URL: http://archimer.ifremer.fr/doc/00302/41336/40522.pdf
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Optimization in Agrobacterium-mediated transformation of Anthurium andraeanum using GFP as a reporter Electron. J. Biotechnol.
Zhao,Qing; Jing,Ji; Wang,Gang; Wang,Jie Hua; Feng,Yuan Yuan; Xing,Han Wen; Guan,Chun Feng.
Although Agrobacterium-mediated transformation protocols for many economically important plant species have been well established, protocol for a number of flowering plants including Anthurium andraeanum remains challenging. In this study, we report success in generating transgenic Anthurium andraeanum cv Arizona using Agrobacterium GV3101 strain harboring a binary vector carrying gfp as a reporter gene. The possibility of facilitating the screening process for transgenic plants expressing functional proteins using gfp marker was explored. In order to realize high transformation efficiency, different explant sources including undifferentiated callus pieces and petioles were compared for their regeneration efficiency and susceptibility to...
Tipo: Journal article Palavras-chave: Cocultivation; Gene transfer; Green fluorescent protein; Regeneration.
Ano: 2010 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500009
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Optimizing expression and purification of an ATP-binding gene gsiA from Escherichia coli k-12 by using GFP fusion Genet. Mol. Biol.
Wang,Zhongshan; Xiang,Quanju; Wang,Guangjun; Wang,Haiyan; Zhang,Yizheng.
The cloning, expression and purification of the glutathione (sulfur) import system ATP-binding protein (gsiA) was carried out. The coding sequence of Escherichia coli gsiA, which encodes the ATP-binding protein of a glutathione importer, was amplified by PCR, and then inserted into a prokaryotic expression vector pWaldo-GFPe harboring green fluorescent protein (GFP) reporter gene. The resulting recombinant plasmid pWaldo-GFP-GsiA was transformed into various E. coli strains, and expression conditions were optimized. The effect of five E. coli expression strains on the production of the recombinant gsiA protein was evaluated. E. coli BL21 (DE3) was found to be the most productive strain for GsiA-GFP fusion-protein expression, most of which was insoluble...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Escherichia coli; Glutathione transporter; GsiA; Gene expression; Green fluorescent protein.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000400019
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Oviduct-specific expression of tissue plasminogen activator in laying hens Genet. Mol. Biol.
Kaleri,Hubdar Ali; Xiang,Liu; Aniwashi,Jueken; Xu,Shiyong.
Egg-laying hens are important candidate bioreactors for pharmaceutical protein production because of the amenability of their eggs for protein expression. In this study, we constructed an oviduct-specific vector containing tissue plasminogen activator (tPA) protein and green fluorescent protein (pL-2.8OVtPAGFP) and assessed its expression in vitro and in vivo. Oviduct epithelial and 3T3 cells were cultured and transfected with pL-2.8OVtPAGFP and pEGP-N1 (control vector), respectively. The pL-2.8OVtPAGFP vector was administered to laying hens via a wing vein and their eggs and tissues were examined for tPA expression. The oviduct-specific vector pL-2.8OVtPAGFP was expressed only in oviduct epithelial cells whereas pEGP-N1 was detected in oviduct epithelial...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Green fluorescent protein; Human tissue plasminogen activator; Laying hens; Oviduct-specific expression.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000200011
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Production of bovine transgenic embryos by microinjection of a lentiviral vector in mature ovocytes. Repositório Alice
OTERO, R.; HERNÁNDEZ, D.; CAMARGO, L. S. de A..
Abstract Objective: To produce bovine transgenic embryos by microinjection of a lentiviral vector with the eGFP gene as a marker. Methods: Four treatments were designed: T1=Control: fertilized in vitro (FIV) with cumulus-oocyte complexes (CCOs), cultivated in CR2 medium with 10% FBS and incubated at 38.5°C in an atmosphere of 95% humidity and 5% CO₂. T2=Control of culture medium: CCOs removed by vortex in the presence of hyaluronidase, FIV, grown in SOF medium in hermetic bag, with a gaseous mixture of 5% CO₂, 5% O₂ and 90% N₂ and humidity saturated at 38.5°C. T3=Microinjection control: CCOs removed microinjected with TALP medium, FIV and cultured under the same treatment conditions T2. T4=Microinjected with the...
Tipo: Artigo em periódico indexado (ALICE) Palavras-chave: Genetic Modification; EGFP; Micromanipulation; Green fluorescent protein.
Ano: 2018 URL: http://www.alice.cnptia.embrapa.br/alice/handle/doc/1102529
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Rat Dlx5 is expressed in the subventricular zone and promotes neuronal differentiation BJMBR
Shu,H.F.; Gao,F.Y.; Zhang,C.Q.; Liu,S.Y.; Zhang,Z.Y.; Song,Y.C.; Qiu,K.J.; Yang,H..
The molecular mechanisms and potential clinical applications of neural precursor cells have recently been the subject of intensive study. Dlx5, a homeobox transcription factor related to the distal-less gene in Drosophila, was shown to play an important role during forebrain development. The subventricular zone (SVZ) in the adult brain harbors the largest abundance of neural precursors. The anterior SVZ (SVZa) contains the most representative neural precursors in the SVZ. Further research is necessary to elucidate how Dlx5-related genes regulate the differentiation of SVZa neural precursors. Here, we employed immunohistochemistry and molecular biology techniques to study the expression of Dlx5 and related homeobox genes Er81 and Islet1 in neonatal rat...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Neural precursors; Dlx5; Green fluorescent protein; Recombinant plasmids.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2010000200008
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Transformation of an Argentine spring wheat genotype: optimization of the protocols for particle bombardment of excised immature embryos and rapid isolation of transgenic plants JBAG
Souza Canada,Eduardo Daniel; Fettig,Sebastian; Ziegler,Paul; Beck,Erwin.
Klein Brujo was found to be a promising Argentine spring wheat genotype for transformation studies. In the present work we optimized the biolistic transformation of embryogenic scutellar calli from this genotype. We first identified scutellar callus induction media (SCIM) most promising for in vitro embryogenic plant regeneration with Klein Brujo. We then co-bombarded embryos of Klein Brujo and, for comparison, of Bobwhite, a highly transformable wheat line, on these media with 1 μm gold particles coated with two plasmids. One of these contained the marker gene gfp (linked to the CaMV35S-promoter) and the selection gene bar (resistance to phosphinothricin: PPT, linked to the maize Ubi1-promoter), whereas the other contained ipt (encoding...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Particle bombardment parameters; Co-transformation; Green fluorescent protein; Phosphinothricin resistance; Transgene segregation.
Ano: 2015 URL: http://www.scielo.org.ar/scielo.php?script=sci_arttext&pid=S1852-62332015000100003
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Use of green fluorescent protein to monitor Lactobacillus plantarum in the gastrointestinal tract of goats BJM
Han,Xufeng; Wang,Lei; Li,Wei; Li,Bibo; Yang,Yuxin; Yan,Hailong; Qu,Lei; Chen,Yulin.
The experiment aimed to specifically monitor the passage of lactobacilli in vivo after oral administration. The green fluorescent protein (GFP) gene was cloned downstream from the constitutive p32 promoter from L. lactis subsp. cremoris Wg2. The recombinant expression vector, pLEM415-gfp-p32, was electroporated into Lactobacillus plantarum (L. plantarum) isolated from goat. Green fluorescent protein (GFP) was successfully expressed in L. plantarum. After 2 h post-administration, transformed Lactobacillus could be detectable in all luminal contents. In the rumen, bacteria concentration initially decreased, reached the minimum at 42 h post-oral administration and then increased. However, this concentration decreased constantly in the duodenum. This result...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Green fluorescent protein; Gastro-intestinal tract; Lactobacillus plantarum; Goat.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000300849
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Using green fluorescent protein to understand the mechanisms of G-protein-coupled receptor regulation BJMBR
Ferguson,S.S.G..
G protein-coupled receptor (GPCR) activation is followed rapidly by adaptive changes that serve to diminish the responsiveness of a cell to further stimulation. This process, termed desensitization, is the consequence of receptor phosphorylation, arrestin binding, sequestration and down-regulation. GPCR phosphorylation is initiated within seconds to minutes of receptor activation and is mediated by both second messenger-dependent protein kinases and receptor-specific G protein-coupled receptor kinases (GRKs). Desensitization in response to GRK-mediated phosphorylation involves the binding of arrestin proteins that serve to sterically uncouple the receptor from its G protein. GPCR sequestration, the endocytosis of receptors to endosomes, not only...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Arrestin; Endocytosis; Green fluorescent protein; G protein-coupled receptors; G protein-coupled receptor kinases; Resensitization.
Ano: 1998 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1998001100016
Registros recuperados: 12
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