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1.
Behav Sci (Basel) ; 14(3)2024 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-38540532

RESUMO

The problems that arise with coexistence between schoolchildren are a challenge when it comes to carrying out the teaching-learning process. In recent years, the presence of bullying has increased in schools. The aim of this study is to identify the perceptions of teachers regarding the problems of coexistence among schoolchildren, more specifically concerning bullying, as well as to identify their position regarding the prevention of bullying. Research was carried out using a mixed approach. A total of 225 education professionals from different schools in Spain took part. From the results and our conclusions, it should be noted that teachers consider themselves to have a high level of knowledge of the essential characteristics of the dynamics of bullying, of the actors involved, and of some preventive measures that can be carried out to avoid this type of behaviour as much as possible. On the other hand, a large number of teachers downplay the importance of the problem of coexistence in their fields, and/or are not able to identify it, which makes it impossible to act. Some schools hardly carry out any prevention work, which makes it difficult to eradicate bullying.

2.
Children (Basel) ; 10(2)2023 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-36832379

RESUMO

Information and Communication Technologies are now a common feature in classroom activities. The aim of this study was to present praxis developed for the tablet for use by primary education students (aged 6-12) studying the natural sciences and mathematics. This research is qualitative and follows the narrative-ethnographic approach. The study sample consisted of 120 primary education students and 52 educational blogs. The results and conclusions reveal praxis that is rarely innovative or ludic. The bulk of tablet-based activities were for natural sciences classes rather than mathematics, and the most common practice with the tablet in the natural sciences was information searching and content exploration. The most widely used apps were the Google search engine, YouTube and the tablet's default apps (camera, image and video editor). Course content in the natural sciences focused on living beings and states of matter, and the activities developed for children to do on the tablet aimed to foster learning through discovery, exploration and enquiry. In mathematics, a traditional methodological approach was apparent in children's use of the tablet for typical activities related to units of measurement.

3.
Artigo em Inglês | MEDLINE | ID: mdl-35162171

RESUMO

Environmental education, at least in northwest Spain, is often overlooked in the education system from infant schooling onwards and interventions are needed to raise the profile of this subject. The aim of this study was to examine the impact of a learning program designed for primary school students to broaden their ecological awareness and improve digital literacy using gamification tools. The research was developed using a qualitative approach, with data obtained from 156 subjects, including teachers, students and families. The results show that the children assimilated new habits on the better usage of water and electricity and recycling paper and plastic. Moreover, they acquired more efficient strategies for finding information online, by using apps and developing content with digital tools. Gaming dynamics and resources were the key to students' learning, with the tablet proving an essential tool for boosting motivation, interaction and problem solving.


Assuntos
Gamificação , Alfabetização , Criança , Humanos , Motivação , Instituições Acadêmicas , Estudantes
4.
Front Microbiol ; 13: 1063706, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36756201

RESUMO

Parvoviruses are promising anticancer and gene therapy agents, but a deep knowledge of the entry process is crucial to exploit their therapeutic potential. We addressed this issue while attempting to retarget the oncolytic parvovirus minute virus of mice (MVMp) to the tumor vasculature. Residues at three functional domains of the icosahedral capsid were substituted by rational design with peptides competing with the vascular endothelial growth factor. Most substitutions impaired virus maturation, though some yielded infectious chimeric virions, and substitutions in a dimple at the twofold axis that allocates sialic acid (SIA) receptors altered viral tropism. One dimple-modified chimeric virion was efficiently attached as MVMp to α2-linked SIA moieties, but the infection was impaired by the binding to some inhibitory α2-3,-6,-8 SIA pseudoreceptors, which hampers intracellular virus traffic to the endosome in a cell type-dependent manner. Infectious from nonproductive traffic could be mechanistically discriminated by an endosomal drastic capsid structural transition comprising the cleavage of some VP2-Nt sequences and its associated VP1-Nt exposure. Correspondingly, neuraminidase removal of inhibitory SIA moieties enhanced the infection quantitatively, correlating to the restored virus traffic to the endosome and the extent of VP2-Nt cleavage/VP1-Nt exposure. This study illustrates (i) structural constraints to retarget parvoviruses with evolutionary adopted narrow grooves allocating small SIA receptors, (ii) the possibility to enhance parvovirus oncolysis by relaxing the glycan network on the cancer cell surface, and (iii) the major role played by the attachment to cell type-specific SIAs in the intracellular virus traffic to the endosome, which may determine parvovirus tropism and host range.

5.
J Virol ; 93(19)2019 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-31315994

RESUMO

As many tumor cells synthetize vascular endothelial growth factors (VEGF) that promote neo-vascularization and metastasis, frontline cancer therapies often administer anti-VEGF (α-VEGF) antibodies. To target the oncolytic parvovirus minute virus of mice (MVM) to the tumor vasculature, we studied the functional tolerance, evasion of neutralization, and induction of α-VEGF antibodies of chimeric viruses in which the footprint of a neutralizing monoclonal antibody within the 3-fold capsid spike was replaced by VEGF-blocking peptides: P6L (PQPRPL) and A7R (ATWLPPR). Both peptides allowed viral genome replication and nuclear translocation of chimeric capsid subunits. MVM-P6L efficiently propagated in culture, exposing the heterologous peptide on the capsid surface, and evaded neutralization by the anti-spike monoclonal antibody. In contrast, MVM-A7R yielded low infectious titers and was poorly recognized by an α-A7R monoclonal antibody. MVM-A7R showed a deficient assembly pattern, suggesting that A7R impaired a transitional configuration that the subunits must undergo in the 3-fold axis to close up the capsid shell. The MVM-A7R chimeric virus consistently evolved in culture into a mutant carrying the P6Q amino acid substitution within the A7R sequence, which restored normal capsid assembly and infectivity. Consistent with this finding, anti-native VEGF antibodies were induced in mice by a single injection of MVM-A7R empty capsids, but not by MVM-A7R virions. This fundamental study provides insights to endow an infectious parvovirus with immune antineovascularization and evasion capacities by replacing an antibody footprint in the capsid 3-fold axis with VEGF-blocking peptides, and it also illustrates the evolutionary capacity of single-stranded DNA (ssDNA) viruses to overcome engineered capsid structural restrictions.IMPORTANCE Targeting the VEGF signaling required for neovascularization by vaccination with chimeric capsids of oncolytic viruses may boost therapy for solid tumors. VEGF-blocking peptides (VEbp) engineered in the capsid 3-fold axis endowed the infectious parvovirus MVM with the ability to induce α-VEGF antibodies without adjuvant and to evade neutralization by MVM-specific antibodies. However, these properties may be compromised by structural restraints that the capsid imposes on the peptide configuration and by misassembly caused by the heterologous peptides. Significantly, chimeric MVM-VEbp resolved the structural restrictions by selecting mutations within the engineered peptides that restored efficient capsid assembly. These data show the promise of antineovascularization vaccines using chimeric VEbp-icosahedral capsids of oncolytic viruses but also raise safety concerns regarding the genetic stability of manipulated infectious parvoviruses in cancer and gene therapies.


Assuntos
Vacinas Anticâncer/imunologia , Proteínas do Capsídeo/imunologia , Proteínas do Capsídeo/metabolismo , Vírus Miúdo do Camundongo/imunologia , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Fator A de Crescimento do Endotélio Vascular/imunologia , Animais , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Vacinas Anticâncer/administração & dosagem , Vacinas Anticâncer/genética , Proteínas do Capsídeo/genética , Camundongos Endogâmicos BALB C , Vírus Miúdo do Camundongo/genética , Vírus Miúdo do Camundongo/crescimento & desenvolvimento , Vírus Oncolíticos/genética , Vírus Oncolíticos/crescimento & desenvolvimento , Vírus Oncolíticos/imunologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Carga Viral , Montagem de Vírus , Ligação Viral , Internalização do Vírus
6.
Virology ; 432(1): 45-56, 2012 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-22727830

RESUMO

The unordered N-termini of parvovirus capsid proteins (Nt) are translocated through a channel at the icosahedral five-fold axis to serve for virus traffick. Heterologous peptides were genetically inserted at the Nt of MVM to study their functional tolerance to manipulations. Insertion of a 5T4-single-chain antibody at VP2-Nt (2Nt) yielded chimeric capsid subunits failing to enter the nucleus. The VEGFR2-binding peptide (V1) inserted at both 2Nt and VP1-Nt efficiently assembled in virions, but V1 disrupted VP1 and VP2 entry functions. The VP2 defect correlated with restricted externalization of V1-2Nt out of the coat. The specific infectivity of MVM and wtVP-pseudotyped mosaic MVM-V1 virions, upon heating and/or partial 2Nt cleavage, demonstrated that some 2Nt domains become intracellularly translocated out of the virus shell and cleaved to initiate entry. The V1 insertion defines a VP2-driven endosomal enlargement of the channel as an essential structural rearrangement performed by the MVM virion to infect.


Assuntos
Proteínas do Capsídeo/metabolismo , Vírus Miúdo do Camundongo/fisiologia , Montagem de Vírus , Internalização do Vírus , Animais , Proteínas do Capsídeo/genética , Linhagem Celular , Núcleo Celular/virologia , Endossomos/virologia , Camundongos , Modelos Biológicos , Estrutura Terciária de Proteína
7.
Neoplasia ; 13(7): 633-42, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21750657

RESUMO

Metastatic melanoma cells are highly adaptable to their in vivo microenvironment and can switch between protease-dependent mesenchymal and protease-independent amoeboid invasion to facilitate metastasis. Such adaptability can be visualized in vitro, when cells are cultured in conditions that recapitulate three-dimensional microenvironments. Using thick collagen layers in cell culture and in vivo extravasation assays, we found that pigment epithelium-derived factor (PEDF) suppressed lung extravasation of aggressive melanoma by coordinated regulation of cell shape and proteolysis. In cells grown on a thick collagen bed, PEDF overexpression and exogenous PEDF blocked the rapidly invasive, rounded morphology, and promoted an elongated, mesenchymal-like phenotype associated with reduced invasion. These changes in cell shape depended on decreased RhoA and increased Rac1 activation and were mediated by the up-regulation of Rac1-GEF, DOCK3 and down-regulation of Rac1-GAP, ARHGAP22. Surprisingly, we found that PEDF overexpression also blocked the trafficking of membrane-tethered, MT1-MMP to the cell surface through RhoA inhibition and Rac1 activation. In vivo, knockdown of Rac1 and DOCK3 or overexpression of MT1-MMP was sufficient to reverse the inhibitory effect of PEDF on extravasation. Using functional studies, we demonstrated that PEDF suppressed the rounded morphology and MT1-MMP surface localization through its antiangiongenic, 34-mer epitope and the recently identified PEDF receptor candidate, PNPLA2. Our findings unveil the coordinated regulation of cell shape and proteolysis and identify an unknown mechanism for PEDF's antimetastatic activity.


Assuntos
Amoeba/citologia , Proteínas do Olho/farmacologia , Mesoderma/efeitos dos fármacos , Invasividade Neoplásica/prevenção & controle , Neoplasias/patologia , Fatores de Crescimento Neural/farmacologia , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Pseudópodes/efeitos dos fármacos , Serpinas/farmacologia , Animais , Antineoplásicos/farmacologia , Forma Celular/efeitos dos fármacos , Células Cultivadas , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Proteínas do Olho/uso terapêutico , Humanos , Mesoderma/metabolismo , Camundongos , Camundongos Nus , Neoplasias/tratamento farmacológico , Fatores de Crescimento Neural/uso terapêutico , Pseudópodes/patologia , Serpinas/uso terapêutico , Ensaios Antitumorais Modelo de Xenoenxerto
8.
Melanoma Res ; 21(4): 285-97, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21673604

RESUMO

Pigment epithelium-derived factor (PEDF) is a broad-spectrum angiogenesis inhibitor that displays potent antimetastatic activity in multiple tumor types. We have previously shown that PEDF prevents primary tumor growth and metastatic spread of human melanoma in mouse experimental models. Consistent with these observations, PEDF expression is lost at the late stages of melanoma progression, allowing melanoma cells to become angiogenic, migratory, and invasive. PEDF's ability to modify the interplay between the host and tumor tissues strongly supports its use as a therapeutic agent for the treatment of metastatic melanoma. However, transition to the clinic requires a more detailed knowledge of the molecular mechanisms underpinning PEDF's activity. In this study, we describe changes in the gene expression profile of A375 human melanoma cells induced by PEDF overexpression. PEDF modulated diverse categories of genes known to be involved in angiogenesis and migration. It downregulated cytokines such as interleukin-8 and extracellular matrix proteins such as collagen IV, while it upregulated fibronectin. Multiple transcripts previously described as contributing to the acquisition of malignant phenotype by melanoma were also diminished by PEDF overexpression, among which we validated galectin 3 and jagged 1. In addition, PEDF downregulated S100ß and melanoma inhibitory activity, which are widely used in the pathological diagnosis of melanoma. Interestingly, PEDF increased the expression of melanophilin and decreased rab27A, which are relevant targets for melanosome transport; suggesting that PEDF could directly impinge on melanocytic lineage-specific processes. Our study identifies new molecular targets and signaling pathways that may potentially contribute to determine PEDF's ability to restrict the aggressiveness of A375 human melanoma cells.


Assuntos
Proteínas do Olho/genética , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Melanoma/genética , Fatores de Crescimento Neural/genética , Serpinas/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proteínas do Olho/metabolismo , Perfilação da Expressão Gênica/métodos , Genótipo , Humanos , Melanoma/metabolismo , Melanoma/patologia , Invasividade Neoplásica/genética , Neovascularização Patológica/genética , Fatores de Crescimento Neural/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Fenótipo , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Serpinas/metabolismo , Transfecção , Regulação para Cima
9.
Blood ; 116(3): 475-84, 2010 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-20203265

RESUMO

Extracellular factors control the angiogenic switch in endothelial cells (ECs) via competing survival and apoptotic pathways. Previously, we showed that proangiogenic and antiangiogenic factors target the same signaling molecules, which thereby become pivots of angiogenic balance. Here we show that in remodeling endothelium (ECs and EC precursors) natural angiogenic inhibitors enhance nuclear factor-kappaB (NF-kappaB) DNA binding, which is critical for antiangiogenesis, and that blocking the NF-kappaB pathway abolishes multiple antiangiogenic events in vitro and in vivo. NF-kappaB induction by antiangiogenic molecules has a dual effect on transcription. NF-kappaB acts as an activator of proapoptotic FasL and as a repressor of prosurvival cFLIP. On the FasL promoter, NF-kappaB increases the recruitment of HAT p300 and acetylated histones H3 and H4. Conversely, on cFLIP promoter, NF-kappaB increases histone deacetylase 1 (HDAC1), decreases p300 and histone acetylation, and reduces the recruitment of NFAT, a transcription factor critical for cFLIP expression. Finally, we found a biphasic effect, when HDAC inhibitors (HDACi) were used to test the dependence of pigment epithelial-derived factor activity on histone acetylation. The cooperative effect seen at low doses switches to antagonistic as the concentrations increase. Our study defines an interactive transcriptional network underlying angiogenic balance and points to HDACi as tools to manipulate the angiogenic switch.


Assuntos
Montagem e Desmontagem da Cromatina/fisiologia , NF-kappa B/fisiologia , Fatores de Transcrição NFATC/fisiologia , Neovascularização Fisiológica , Inibidores da Angiogênese/farmacologia , Animais , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/genética , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Proteínas do Olho/farmacologia , Proteína Ligante Fas/genética , Proteína Ligante Fas/metabolismo , Feminino , Histona Desacetilase 1/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Nus , Fatores de Transcrição NFATC/genética , Neovascularização Fisiológica/efeitos dos fármacos , Neovascularização Fisiológica/genética , Fatores de Crescimento Neural/farmacologia , Regiões Promotoras Genéticas , Serpinas/farmacologia , Transdução de Sinais , Trombospondina 1/farmacologia
10.
FEBS Lett ; 581(16): 3033-8, 2007 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-17544406

RESUMO

Two phosphofructokinase (PFK) chimeras were constructed by exchanging the N- and C-terminal halves of the mammalian M- and C-type isozymes, to investigate the contribution of each terminus to the catalytic site and the fructose-2,6-P(2)/fructose-1,6-P(2) allosteric site. The homogeneously-purified chimeric enzymes organized into tetramers, and exhibited kinetic properties for fructose-6-P and MgATP similar to those of the native enzyme that furnished the N-terminal domain in each case, whereas their fructose-2,6-P(2) activatory characteristics coincided with those of the isozyme that provided the C-terminal half. This reflected the role of each domain in the formation of the corresponding binding site. Grafting the N-terminus of PFK-M onto the C-terminus of the fructose-1,6-P(2) insensitive PFK-C restored transduction of this signal to the catalytic site, which significance is also discussed.


Assuntos
Frutosedifosfatos/metabolismo , Fosfofrutoquinases/química , Fosfofrutoquinases/metabolismo , Trifosfato de Adenosina/metabolismo , Sítios de Ligação , Humanos , Isoenzimas/química , Isoenzimas/metabolismo , Ligantes , Fosfofrutoquinase-1 Muscular/química , Fosfofrutoquinase-1 Muscular/metabolismo , Fosfofrutoquinase-1 Tipo C/química , Fosfofrutoquinase-1 Tipo C/metabolismo , Estrutura Terciária de Proteína , Proteínas Recombinantes de Fusão/síntese química , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/metabolismo , Saccharomyces cerevisiae
11.
J Biol Chem ; 280(18): 17969-77, 2005 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-15728575

RESUMO

In the spherical virion of the parvovirus minute virus of mice, several amino acid side chains of the capsid were previously found to be involved in interactions with the viral single-stranded DNA molecule. We have individually truncated by mutation to alanine many (ten) of these side chains and analyzed the effects on capsid assembly, stability and conformation, viral DNA encapsidation, and virion infectivity. Mutation of residues Tyr-270, Asp-273, or Asp-474 led to a drastic reduction in infectivity. Mutant Y270A was defective in capsid assembly; mutant D273A formed stable capsids, but it was essentially unable to encapsidate the viral DNA or to externalize the N terminus of the capsid protein VP2, a connected conformational event. Mutation of residues Asp-58, Trp-60, Asn-183, Thr-267, or Lys-471 led to a moderate reduction in infectivity. None of these mutations had an effect on capsid assembly or stability, or on the DNA encapsidation process. However, those five mutant virions were substantially less stable than the parental virion in thermal inactivation assays. The results with this model spherical virus indicate that several capsid residues that are found to be involved in polar interactions or multiple hydrophobic contacts with the viral DNA molecule contribute to preserving the active conformation of the infectious viral particle. Their effect appears to be mediated by the non-covalent interactions they establish with the viral DNA. In addition, at least one acidic residue at each DNA-binding region is needed for DNA packaging.


Assuntos
Parvovirus/química , Parvovirus/genética , Vírion/química , Vírion/genética , Aminoácidos/química , Aminoácidos/genética , Animais , Proteínas do Capsídeo/química , Proteínas do Capsídeo/genética , Linhagem Celular , Análise Mutacional de DNA/métodos , Camundongos , Mutagênese Sítio-Dirigida , Ácidos Nucleicos/química , Ácidos Nucleicos/genética
12.
Biochem J ; 377(Pt 1): 77-84, 2004 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-12974670

RESUMO

Systematic deletions and point mutations in the C-terminal extension of mammalian PFK (phosphofructokinase) led us to identify Leu-767 and Glu-768 of the M-type isoform (PFK-M) as the motifs responsible for the role of this region in inhibition by MgATP. These amino acids are the only residues of the C-terminus that are conserved in all mammalian isoforms, and were found to have a similar function in the C-type isoenzyme. Both residues in PFK-C and Leu-767 in PFK-M were also observed to be critical for inhibition by citrate, which is synergistic with that by MgATP. Binding studies utilizing titration of intrinsic protein fluorescence indicated that the C-terminal part of the enzyme participates in the signal transduction route from the MgATP inhibitory site to the catalytic site, but does not contribute to the binding of this inhibitor, whereas it is essential for the binding of citrate. Mutations of the identified structural motifs did not alter either the action of other allosteric effectors that also interact with MgATP, such as the inhibitor phosphoenolpyruvate and the strong activator fructose 2,6-bisphosphate, or the co-operative effect of fructose 6-phosphate. The latter data provide evidence that activation by fructose 2,6-bisphosphate and fructose 6-phosphate co-operativity are not linked to the same allosteric transition as that mediating inhibition by MgATP.


Assuntos
Trifosfato de Adenosina/farmacologia , Mamíferos/metabolismo , Fosfofrutoquinases/química , Fosfofrutoquinases/metabolismo , Trifosfato de Adenosina/metabolismo , Regulação Alostérica , Motivos de Aminoácidos , Sequência de Aminoácidos , Animais , Ácido Cítrico/metabolismo , Ácido Cítrico/farmacologia , Histidina/fisiologia , Humanos , Isoenzimas/química , Isoenzimas/metabolismo , Dados de Sequência Molecular , Mutação , Fosfofrutoquinases/genética , Alinhamento de Sequência
13.
Eukaryot Cell ; 1(6): 865-74, 2002 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-12477787

RESUMO

Candida albicans is able to respond to environmental changes by inducing a distinct morphological program, which is related to the ability to infect mammalian hosts. Although some of the signal transduction pathways involved in this response are known, it is not clear how the environmental signals are sensed and transmitted to these transduction cascades. In this work, we have studied the function of GPA2, a new gene from C. albicans, which encodes a G-protein alpha-subunit homologue. We demonstrate that Gpa2 plays an important role in the yeast-hypha dimorphic transition in the response of C. albicans to some environmental inducers. Deletion of both alleles of the GPA2 gene causes in vitro defects in morphological transitions in Spider medium and SLAD medium and in embedded conditions but not in medium containing serum. These defects cannot be reversed by exogenous addition of cyclic AMP. However, overexpression of HST7, which encodes a component of the filament-inducing mitogen-activated protein kinase (MAPK) cascade, bypasses the Gpa2 requirement. We have obtained different gain-of-function and loss-of-function mutant alleles of the GPA2 gene, which we have introduced in several C. albicans genetic backgrounds. Our results indicate that, in response to environmental cues, Gpa2 is required for the regulation of a MAPK signaling pathway.


Assuntos
Candida albicans/fisiologia , Proteínas Fúngicas/química , Proteínas Fúngicas/fisiologia , Subunidades alfa de Proteínas de Ligação ao GTP , Proteínas Heterotriméricas de Ligação ao GTP/química , Proteínas Heterotriméricas de Ligação ao GTP/fisiologia , Hifas/fisiologia , Alelos , Sequência de Aminoácidos , Southern Blotting , AMP Cíclico/metabolismo , Proteínas Fúngicas/genética , Proteínas Heterotriméricas de Ligação ao GTP/genética , Sistema de Sinalização das MAP Quinases , Modelos Genéticos , Dados de Sequência Molecular , Mutação , Plasmídeos/metabolismo , Proteínas de Saccharomyces cerevisiae , Homologia de Sequência de Aminoácidos , Transdução de Sinais , Temperatura , Fatores de Tempo , Transgenes
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