Base de dados : MEDLINE
Pesquisa : A11.284.187.178.170 [Categoria DeCS]
Referências encontradas : 3985 [refinar]
Mostrando: 1 .. 10   no formato [Detalhado]

página 1 de 399 ir para página                         

  1 / 3985 MEDLINE  
              next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:29237735
[Au] Autor:Dougherty JD
[Ad] Endereço:Departments of Genetics and Psychiatry, Washington University School of Medicine, St. Louis, Missouri 63110 jdougherty@genetics.wustl.edu.
[Ti] Título:The Expanding Toolkit of Translating Ribosome Affinity Purification.
[So] Source:J Neurosci;37(50):12079-12087, 2017 Dec 13.
[Is] ISSN:1529-2401
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Translating ribosome affinity purification is a method initially developed for profiling mRNA from genetically defined cell types in complex tissues. It has been applied both to identify target molecules in cell types that are important for controlling a variety of behaviors in the brain, and to understand the molecular consequences on those cells due to experimental manipulations, ranging from drugs of abuse to disease-causing mutations. Since its inception, a variety of methodological advances are opening new avenues of investigation. These advances include a variety of new methods for targeting cells for translating ribosome affinity purification by features such as their projections or activity, additional tags and mouse reagents increasing the flexibility of the system, and new modifications of the method specifically focused on studying the regulation of translation. The latter includes methods to assess cell type-specific regulation of translation in specific subcellular compartments. Here, I provide a summary of these recent advances and resources, highlighting both new experimental opportunities and areas for future technical development.
[Mh] Termos MeSH primário: Fracionamento Celular/métodos
Perfilação da Expressão Gênica/métodos
Ensaios de Triagem em Larga Escala/métodos
Separação Imunomagnética/métodos
Neuroglia/ultraestrutura
Neurônios/ultraestrutura
Biossíntese de Proteínas
Ribossomos
[Mh] Termos MeSH secundário: Marcadores de Afinidade
Animais
Encéfalo/citologia
Fracionamento Celular/tendências
Linhagem Celular
Cromossomos Artificiais Bacterianos
Dependovirus/genética
Eletroporação
Imunofluorescência
Previsões
Genes Reporter
Vetores Genéticos/genética
Proteínas de Fluorescência Verde/análise
Proteínas de Fluorescência Verde/genética
Camundongos
Neuroglia/metabolismo
Neurônios/classificação
Neurônios/metabolismo
Fases de Leitura Aberta/genética
RNA Mensageiro/genética
RNA Mensageiro/isolamento & purificação
Proteínas Recombinantes de Fusão/análise
Proteínas Ribossômicas/análise
Proteínas Ribossômicas/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Affinity Labels); 0 (RNA, Messenger); 0 (Recombinant Fusion Proteins); 0 (Ribosomal Proteins); 147336-22-9 (Green Fluorescent Proteins)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180130
[Lr] Data última revisão:
180130
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171215
[St] Status:MEDLINE
[do] DOI:10.1523/JNEUROSCI.1929-17.2017


  2 / 3985 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:28743798
[Au] Autor:Chen W; Huang H; Hatori R; Kornberg TB
[Ad] Endereço:Cardiovascular Research Institute, University of California, San Francisco, CA 94143, USA.
[Ti] Título:Essential basal cytonemes take up Hedgehog in the wing imaginal disc.
[So] Source:Development;144(17):3134-3144, 2017 09 01.
[Is] ISSN:1477-9129
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Morphogen concentration gradients that extend across developmental fields form by dispersion from source cells. In the wing disc, Hedgehog (Hh) produced by posterior compartment cells distributes in a concentration gradient to adjacent cells of the anterior compartment. We monitored Hh:GFP after pulsed expression, and analyzed the movement and colocalization of Hh, Patched (Ptc) and Smoothened (Smo) proteins tagged with GFP or mCherry and expressed at physiological levels from bacterial artificial chromosome transgenes. Hh:GFP moved to basal subcellular locations prior to release from posterior compartment cells that express it, and was taken up by basal cytonemes that extend to the source cells. Hh and Ptc were present in puncta that moved along the basal cytonemes and formed characteristic apical-basal distributions in the anterior compartment cells. The basal cytonemes required , , and , and both the Hh gradient and Hh signaling declined under conditions in which the cytonemes were compromised. These findings show that in the wing disc, Hh distributions and signaling are dependent upon basal release and uptake, and on cytoneme-mediated movement. No evidence for apical dispersion was obtained.
[Mh] Termos MeSH primário: Proteínas de Drosophila/metabolismo
Drosophila melanogaster/metabolismo
Proteínas Hedgehog/metabolismo
Discos Imaginais/metabolismo
Asas de Animais/metabolismo
[Mh] Termos MeSH secundário: Animais
Padronização Corporal
Compartimento Celular
Cromossomos Artificiais Bacterianos/genética
Proteínas de Fluorescência Verde/metabolismo
Transporte Proteico
Transdução de Sinais
Transgenes
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL
[Nm] Nome de substância:
0 (Drosophila Proteins); 0 (Hedgehog Proteins); 147336-22-9 (Green Fluorescent Proteins); 149291-21-4 (hedgehog protein, Drosophila)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:180112
[Lr] Data última revisão:
180112
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170727
[St] Status:MEDLINE
[do] DOI:10.1242/dev.149856


  3 / 3985 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:27775877
[Au] Autor:Muñoz-Amatriaín M; Mirebrahim H; Xu P; Wanamaker SI; Luo M; Alhakami H; Alpert M; Atokple I; Batieno BJ; Boukar O; Bozdag S; Cisse N; Drabo I; Ehlers JD; Farmer A; Fatokun C; Gu YQ; Guo YN; Huynh BL; Jackson SA; Kusi F; Lawley CT; Lucas MR; Ma Y; Timko MP; Wu J; You F; Barkley NA; Roberts PA; Lonardi S; Close TJ
[Ad] Endereço:Department of Botany and Plant Sciences, University of California, Riverside, CA, USA.
[Ti] Título:Genome resources for climate-resilient cowpea, an essential crop for food security.
[So] Source:Plant J;89(5):1042-1054, 2017 Mar.
[Is] ISSN:1365-313X
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Cowpea (Vigna unguiculata L. Walp.) is a legume crop that is resilient to hot and drought-prone climates, and a primary source of protein in sub-Saharan Africa and other parts of the developing world. However, genome resources for cowpea have lagged behind most other major crops. Here we describe foundational genome resources and their application to the analysis of germplasm currently in use in West African breeding programs. Resources developed from the African cultivar IT97K-499-35 include a whole-genome shotgun (WGS) assembly, a bacterial artificial chromosome (BAC) physical map, and assembled sequences from 4355 BACs. These resources and WGS sequences of an additional 36 diverse cowpea accessions supported the development of a genotyping assay for 51 128 SNPs, which was then applied to five bi-parental RIL populations to produce a consensus genetic map containing 37 372 SNPs. This genetic map enabled the anchoring of 100 Mb of WGS and 420 Mb of BAC sequences, an exploration of genetic diversity along each linkage group, and clarification of macrosynteny between cowpea and common bean. The SNP assay enabled a diversity analysis of materials from West African breeding programs. Two major subpopulations exist within those materials, one of which has significant parentage from South and East Africa and more diversity. There are genomic regions of high differentiation between subpopulations, one of which coincides with a cluster of nodulin genes. The new resources and knowledge help to define goals and accelerate the breeding of improved varieties to address food security issues related to limited-input small-holder farming and climate stress.
[Mh] Termos MeSH primário: Produtos Agrícolas/genética
Produtos Agrícolas/fisiologia
Vigna/genética
Vigna/fisiologia
[Mh] Termos MeSH secundário: Cromossomos Artificiais Bacterianos
Cromossomos de Plantas/genética
Clima
Abastecimento de Alimentos
Genoma de Planta/genética
Genótipo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171128
[Lr] Data última revisão:
171128
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161025
[St] Status:MEDLINE
[do] DOI:10.1111/tpj.13404


  4 / 3985 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:28821183
[Au] Autor:Peichel CL; Sullivan ST; Liachko I; White MA
[Ad] Endereço:Divisons of Basic Sciences and Human Biology, Fred Hutchinson Cancer Research Center, Seattle, WA
[Ti] Título:Improvement of the Threespine Stickleback Genome Using a Hi-C-Based Proximity-Guided Assembly.
[So] Source:J Hered;108(6):693-700, 2017 09 01.
[Is] ISSN:1465-7333
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Scaffolding genomes into complete chromosome assemblies remains challenging even with the rapidly increasing sequence coverage generated by current next-generation sequence technologies. Even with scaffolding information, many genome assemblies remain incomplete. The genome of the threespine stickleback (Gasterosteus aculeatus), a fish model system in evolutionary genetics and genomics, is not completely assembled despite scaffolding with high-density linkage maps. Here, we first test the ability of a Hi-C based proximity-guided assembly (PGA) to perform a de novo genome assembly from relatively short contigs. Using Hi-C based PGA, we generated complete chromosome assemblies from a distribution of short contigs (20-100 kb). We found that 96.40% of contigs were correctly assigned to linkage groups (LGs), with ordering nearly identical to the previous genome assembly. Using available bacterial artificial chromosome (BAC) end sequences, we provide evidence that some of the few discrepancies between the Hi-C assembly and the existing assembly are due to structural variation between the populations used for the 2 assemblies or errors in the existing assembly. This Hi-C assembly also allowed us to improve the existing assembly, assigning over 60% (13.35 Mb) of the previously unassigned (~21.7 Mb) contigs to LGs. Together, our results highlight the potential of the Hi-C based PGA method to be used in combination with short read data to perform relatively inexpensive de novo genome assemblies. This approach will be particularly useful in organisms in which it is difficult to perform linkage mapping or to obtain high molecular weight DNA required for other scaffolding methods.
[Mh] Termos MeSH primário: Mapeamento Cromossômico/métodos
Smegmamorpha/genética
[Mh] Termos MeSH secundário: Alaska
Animais
Cromossomos Artificiais Bacterianos
Mapeamento de Sequências Contíguas
Genômica
Masculino
Análise de Sequência de DNA/métodos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170914
[Lr] Data última revisão:
170914
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170820
[St] Status:MEDLINE
[do] DOI:10.1093/jhered/esx058


  5 / 3985 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:28692067
[Au] Autor:Baumgart L; Mather W; Hasty J
[Ad] Endereço:Molecular Biology Section, Division of Biological Science, University of California, San Diego, La Jolla, California, USA.
[Ti] Título:Synchronized DNA cycling across a bacterial population.
[So] Source:Nat Genet;49(8):1282-1285, 2017 Aug.
[Is] ISSN:1546-1718
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:A defining goal of synthetic biology is to engineer cells to coordinate tasks that often require precise temporal modulation of gene expression. Although a variety of relatively small gene circuits have been constructed and characterized, their logical combination into larger networks remains a central challenge. This is due primarily to the lack of compatible and orthogonal elements for predictable dynamic control of gene expression. As an alternative approach to promoter-level regulation, we explored the use of DNA copy number as a circuit control element. We engineered colony-wide DNA cycling in Escherichia coli in the form of plasmid copy number oscillations via a modular design that can be readily adapted for use with other gene circuitry. Copy number modulation is a generalizable principle that adds a layer of control to synthetic gene circuits, allowing dynamic regulation of circuit elements without requiring specially engineered promoters.
[Mh] Termos MeSH primário: DNA Bacteriano/metabolismo
Escherichia coli/genética
[Mh] Termos MeSH secundário: Cromossomos Artificiais Bacterianos
Variações do Número de Cópias de DNA
Regulação Bacteriana da Expressão Gênica
Redes Reguladoras de Genes
Engenharia Genética
Plasmídeos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Bacterial)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170921
[Lr] Data última revisão:
170921
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170711
[St] Status:MEDLINE
[do] DOI:10.1038/ng.3915


  6 / 3985 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:28671685
[Au] Autor:Capellini TD; Chen H; Cao J; Doxey AC; Kiapour AM; Schoor M; Kingsley DM
[Ad] Endereço:Human Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA.
[Ti] Título:Ancient selection for derived alleles at a GDF5 enhancer influencing human growth and osteoarthritis risk.
[So] Source:Nat Genet;49(8):1202-1210, 2017 Aug.
[Is] ISSN:1546-1718
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Variants in GDF5 are associated with human arthritis and decreased height, but the causal mutations are still unknown. We surveyed the Gdf5 locus for regulatory regions in transgenic mice and fine-mapped separate enhancers controlling expression in joints versus growing ends of long bones. A large downstream regulatory region contains a novel growth enhancer (GROW1), which is required for normal Gdf5 expression at ends of developing bones and for normal bone lengths in vivo. Human GROW1 contains a common base-pair change that decreases enhancer activity and colocalizes with peaks of positive selection in humans. The derived allele is rare in Africa but common in Eurasia and is found in Neandertals and Denisovans. Our results suggest that an ancient regulatory variant in GROW1 has been repeatedly selected in northern environments and that past selection on growth phenotypes explains the high frequency of a GDF5 haplotype that also increases arthritis susceptibility in many human populations.
[Mh] Termos MeSH primário: Elementos Facilitadores Genéticos
Fator 5 de Diferenciação de Crescimento/genética
Osteoartrite/genética
Seleção Genética
[Mh] Termos MeSH secundário: Alelos
Animais
Cromossomos Artificiais Bacterianos
Evolução Molecular
Feminino
Predisposição Genética para Doença
Variação Genética
Seres Humanos
Masculino
Camundongos
Camundongos Endogâmicos C57BL
Camundongos Endogâmicos CBA
Camundongos Transgênicos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (GDF5 protein, human); 0 (Growth Differentiation Factor 5)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170921
[Lr] Data última revisão:
170921
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170704
[St] Status:MEDLINE
[do] DOI:10.1038/ng.3911


  7 / 3985 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:28648842
[Au] Autor:Gallagher TL; Tietz KT; Morrow ZT; McCammon JM; Goldrich ML; Derr NL; Amacher SL
[Ad] Endereço:Molecular Genetics, The Ohio State University, Columbus, OH 43210, United States; Center for RNA Biology, The Ohio State University, Columbus, OH 43210, United States.
[Ti] Título:Pnrc2 regulates 3'UTR-mediated decay of segmentation clock-associated transcripts during zebrafish segmentation.
[So] Source:Dev Biol;429(1):225-239, 2017 09 01.
[Is] ISSN:1095-564X
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Vertebrate segmentation is controlled by the segmentation clock, a molecular oscillator that regulates gene expression and cycles rapidly. The expression of many genes oscillates during segmentation, including hairy/Enhancer of split-related (her or Hes) genes, which encode transcriptional repressors that auto-inhibit their own expression, and deltaC (dlc), which encodes a Notch ligand. We previously identified the tortuga (tor) locus in a zebrafish forward genetic screen for genes involved in cyclic transcript regulation and showed that cyclic transcripts accumulate post-splicing in tor mutants. Here we show that cyclic mRNA accumulation in tor mutants is due to loss of pnrc2, which encodes a proline-rich nuclear receptor co-activator implicated in mRNA decay. Using an inducible in vivo reporter system to analyze transcript stability, we find that the her1 3'UTR confers Pnrc2-dependent instability to a heterologous transcript. her1 mRNA decay is Dicer-independent and likely employs a Pnrc2-Upf1-containing mRNA decay complex. Surprisingly, despite accumulation of cyclic transcripts in pnrc2-deficient embryos, we find that cyclic protein is expressed normally. Overall, we show that Pnrc2 promotes 3'UTR-mediated decay of developmentally-regulated segmentation clock transcripts and we uncover an additional post-transcriptional regulatory layer that ensures oscillatory protein expression in the absence of cyclic mRNA decay.
[Mh] Termos MeSH primário: Regiões 3´ não Traduzidas/genética
Relógios Biológicos/genética
Padronização Corporal/genética
Transativadores/metabolismo
Proteínas de Peixe-Zebra/metabolismo
Peixe-Zebra/embriologia
Peixe-Zebra/genética
[Mh] Termos MeSH secundário: Alelos
Animais
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética
Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo
Cromossomos/genética
Cromossomos Artificiais Bacterianos/genética
Embrião não Mamífero/metabolismo
Regulação da Expressão Gênica no Desenvolvimento
Genes Reporter
Mutação/genética
Degradação do RNAm Mediada por Códon sem Sentido/genética
Fenótipo
Estabilidade de RNA/genética
RNA Mensageiro/genética
RNA Mensageiro/metabolismo
Transativadores/genética
Proteínas de Peixe-Zebra/genética
Zigoto/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (3' Untranslated Regions); 0 (Basic Helix-Loop-Helix Transcription Factors); 0 (Pnrc2 protein, zebrafish); 0 (RNA, Messenger); 0 (Trans-Activators); 0 (Zebrafish Proteins); 0 (her1 protein, zebrafish)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171111
[Lr] Data última revisão:
171111
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170627
[St] Status:MEDLINE


  8 / 3985 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:28582546
[Au] Autor:Baker O; Tsurkan S; Fu J; Klink B; Rump A; Obst M; Kranz A; Schröck E; Anastassiadis K; Stewart AF
[Ad] Endereço:Stem Cell Engineering, Biotechnology Center, Technische Universität Dresden, BioInnovationsZentrum, Tatzberg 47, Dresden 01307, Germany.
[Ti] Título:The contribution of homology arms to nuclease-assisted genome engineering.
[So] Source:Nucleic Acids Res;45(13):8105-8115, 2017 Jul 27.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Designer nucleases like CRISPR/Cas9 enable fluent site-directed damage or small mutations in many genomes. Strategies for their use to achieve more complex tasks like regional exchanges for gene humanization or the establishment of conditional alleles are still emerging. To optimize Cas9-assisted targeting, we measured the relationship between targeting frequency and homology length in targeting constructs using a hypoxanthine-guanine phosphoribosyl-transferase assay in mouse embryonic stem cells. Targeting frequency with supercoiled plasmids improved steeply up to 2 kb total homology and continued to increase with even longer homology arms, thereby implying that Cas9-assisted targeting efficiencies can be improved using homology arms of 1 kb or greater. To humanize the Kmt2d gene, we built a hybrid mouse/human targeting construct in a bacterial artificial chromosome by recombineering. To simplify the possible outcomes, we employed a single Cas9 cleavage strategy and best achieved the intended 42 kb regional exchange with a targeting construct including a very long homology arm to recombine ∼42 kb away from the cleavage site. We recommend the use of long homology arm targeting constructs for accurate and efficient complex genome engineering, particularly when combined with the simplifying advantages of using just one Cas9 cleavage at the genome target site.
[Mh] Termos MeSH primário: Sistemas CRISPR-Cas
Engenharia Genética/métodos
[Mh] Termos MeSH secundário: Animais
Cromossomos Artificiais Bacterianos/genética
Proteínas de Ligação a DNA/genética
Células-Tronco Embrionárias/metabolismo
Endonucleases/metabolismo
Marcação de Genes
Seres Humanos
Hibridização Genética
Hipoxantina Fosforribosiltransferase/genética
Camundongos
Mutação
Proteína de Leucina Linfoide-Mieloide/genética
Proteínas de Neoplasias/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA-Binding Proteins); 0 (MLL2 protein, human); 0 (Mll2 protein, mouse); 0 (Neoplasm Proteins); 149025-06-9 (Myeloid-Lymphoid Leukemia Protein); EC 2.4.2.8 (Hypoxanthine Phosphoribosyltransferase); EC 3.1.- (Endonucleases)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171010
[Lr] Data última revisão:
171010
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170606
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkx497


  9 / 3985 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:28569150
[Au] Autor:Pyeon HR; Nah HJ; Kang SH; Choi SS; Kim ES
[Ad] Endereço:Department of Biological Engineering, Inha University, Incheon, 402-751, South Korea.
[Ti] Título:Heterologous expression of pikromycin biosynthetic gene cluster using Streptomyces artificial chromosome system.
[So] Source:Microb Cell Fact;16(1):96, 2017 May 31.
[Is] ISSN:1475-2859
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Heterologous expression of biosynthetic gene clusters of natural microbial products has become an essential strategy for titer improvement and pathway engineering of various potentially-valuable natural products. A Streptomyces artificial chromosomal conjugation vector, pSBAC, was previously successfully applied for precise cloning and tandem integration of a large polyketide tautomycetin (TMC) biosynthetic gene cluster (Nah et al. in Microb Cell Fact 14(1):1, 2015), implying that this strategy could be employed to develop a custom overexpression scheme of natural product pathway clusters present in actinomycetes. RESULTS: To validate the pSBAC system as a generally-applicable heterologous overexpression system for a large-sized polyketide biosynthetic gene cluster in Streptomyces, another model polyketide compound, the pikromycin biosynthetic gene cluster, was preciously cloned and heterologously expressed using the pSBAC system. A unique HindIII restriction site was precisely inserted at one of the border regions of the pikromycin biosynthetic gene cluster within the chromosome of Streptomyces venezuelae, followed by site-specific recombination of pSBAC into the flanking region of the pikromycin gene cluster. Unlike the previous cloning process, one HindIII site integration step was skipped through pSBAC modification. pPik001, a pSBAC containing the pikromycin biosynthetic gene cluster, was directly introduced into two heterologous hosts, Streptomyces lividans and Streptomyces coelicolor, resulting in the production of 10-deoxymethynolide, a major pikromycin derivative. When two entire pikromycin biosynthetic gene clusters were tandemly introduced into the S. lividans chromosome, overproduction of 10-deoxymethynolide and the presence of pikromycin, which was previously not detected, were both confirmed. Moreover, comparative qRT-PCR results confirmed that the transcription of pikromycin biosynthetic genes was significantly upregulated in S. lividans containing tandem clusters of pikromycin biosynthetic gene clusters. CONCLUSIONS: The 60 kb pikromycin biosynthetic gene cluster was isolated in a single integration pSBAC vector. Introduction of the pikromycin biosynthetic gene cluster into the pikromycin non-producing strains resulted in higher pikromycin production. The utility of the pSBAC system as a precise cloning tool for large-sized biosynthetic gene clusters was verified through heterologous expression of the pikromycin biosynthetic gene cluster. Moreover, this pSBAC-driven heterologous expression strategy was confirmed to be an ideal approach for production of low and inconsistent natural products such as pikromycin in S. venezuelae, implying that this strategy could be employed for development of a custom overexpression scheme of natural product biosynthetic gene clusters in actinomycetes.
[Mh] Termos MeSH primário: Cromossomos Artificiais Bacterianos/genética
Macrolídeos/metabolismo
Família Multigênica/genética
Streptomyces/genética
[Mh] Termos MeSH secundário: Clonagem Molecular
Streptomyces/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Macrolides); FBM8G3Z439 (picromycin)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171108
[Lr] Data última revisão:
171108
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170602
[St] Status:MEDLINE
[do] DOI:10.1186/s12934-017-0708-7


  10 / 3985 MEDLINE  
              first record previous record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:28564645
[Au] Autor:Galkina S; Fillon V; Saifitdinova A; Daks A; Kulak M; Dyomin A; Koshel E; Gaginskaya ER
[Ad] Endereço:Biological Faculty, Saint Petersburg State University, Saint Petersburg, Russia.
[Ti] Título:Chicken Microchromosomes in the Lampbrush Phase: A Cytogenetic Description.
[So] Source:Cytogenet Genome Res;152(1):46-54, 2017.
[Is] ISSN:1424-859X
[Cp] País de publicação:Switzerland
[La] Idioma:eng
[Ab] Resumo:Lampbrush chromosomes are giant, transcriptionally active, meiotic chromosomes found in oocytes of all vertebrates with the exception of mammals. Lampbrush chromosomes offer a convenient tool for cytogenetic mapping and, in particular, have been instrumental in mapping genes and linkage groups on chicken (GGA) chromosomes. Whereas cytogenetic maps of macrochromosome GGA1-10 and microchromosome GGA11-16 lampbrush bivalents have been established, identification and description of smaller microchromosome bivalents are still missing. In this work, we used specific FISH probes for the identification of 12 chicken lampbrush chromosomes formed by GGA17-28. Our observations on chromomere and lateral loop arrangement and chiasma position allowed us to construct the respective cytogenetic maps for these microchromosomes. For the 10 smallest chicken microchromosomes, GGA29-38, no individual molecular tags are available, yet they can be collectively marked using the PO41 repeat. The reported results contribute to building of working cytogenetic maps of the chicken karyotype.
[Mh] Termos MeSH primário: Galinhas/genética
Cromossomos/genética
Citogenética/métodos
[Mh] Termos MeSH secundário: Animais
Cromossomos Artificiais Bacterianos/genética
Metáfase/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170929
[Lr] Data última revisão:
170929
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170601
[St] Status:MEDLINE
[do] DOI:10.1159/000475563



página 1 de 399 ir para página                         
   


Refinar a pesquisa
  Base de dados : MEDLINE Formulário avançado   

    Pesquisar no campo  
1  
2
3
 
           



Search engine: iAH v2.6 powered by WWWISIS

BIREME/OPAS/OMS - Centro Latino-Americano e do Caribe de Informação em Ciências da Saúde