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Pesquisa : B04.280.092.150 [Categoria DeCS]
Referências encontradas : 612 [refinar]
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[PMID]:29327227
[Au] Autor:Khan A; Shrestha A; Bhuyan K; Maiti IB; Dey N
[Ad] Endereço:Department of Gene Function and Regulation, Department of Biotechnology, Government of India, Institute of Life Sciences, Chandrasekharpur, Bhubaneswar, Odisha, India.
[Ti] Título:Structural characterization of a novel full-length transcript promoter from Horseradish Latent Virus (HRLV) and its transcriptional regulation by multiple stress responsive transcription factors.
[So] Source:Plant Mol Biol;96(1-2):179-196, 2018 Jan.
[Is] ISSN:1573-5028
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:KEY MESSAGE: The promoter fragment described in this study can be employed for strong transgene expression under both biotic and abiotic stress conditions. Plant-infecting Caulimoviruses have evolved multiple regulatory mechanisms to address various environmental stimuli during the course of evolution. One such mechanism involves the retention of discrete stress responsive cis-elements which are required for their survival and host-specificity. Here we describe the characterization of a novel Caulimoviral promoter isolated from Horseradish Latent Virus (HRLV) and its regulation by multiple stress responsive Transcription factors (TFs) namely DREB1, AREB1 and TGA1a. The activity of full length transcript (Flt-) promoter from HRLV (- 677 to + 283) was investigated in both transient and transgenic assays where we identified H12 (- 427 to + 73) as the highest expressing fragment having ~ 2.5-fold stronger activity than the CaMV35S promoter. The H12 promoter was highly active and near-constitutive in the vegetative and reproductive parts of both Tobacco and Arabidopsis transgenic plants. Interestingly, H12 contains a distinct cluster of cis-elements like dehydration-responsive element (DRE-core; GCCGAC), an ABA-responsive element (ABRE; ACGTGTC) and as-1 element (TGACG) which are known to be induced by cold, drought and pathogen/SA respectively. The specific binding of DREB1, AREB1 and TGA1a to DRE, ABRE and as-1 elements respectively were confirmed by the gel-binding assays using H12 promoter-specific probes. Detailed mutational analysis of the H12 promoter suggested that the presence of DRE-core and as-1 element was indispensable for its activity which was further confirmed by the transactivation assays. Our studies imply that H12 could be a valuable genetic tool for regulated transgene expression under diverse environmental conditions.
[Mh] Termos MeSH primário: Armoracia/metabolismo
Armoracia/virologia
Caulimovirus/genética
Caulimovirus/metabolismo
Regiões Promotoras Genéticas/genética
Fatores de Transcrição/genética
Fatores de Transcrição/metabolismo
[Mh] Termos MeSH secundário: Arabidopsis/genética
Arabidopsis/metabolismo
Arabidopsis/virologia
Armoracia/genética
Regulação da Expressão Gênica de Plantas
Plantas Geneticamente Modificadas/genética
Plantas Geneticamente Modificadas/metabolismo
Plantas Geneticamente Modificadas/virologia
Tabaco/genética
Tabaco/metabolismo
Tabaco/virologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Transcription Factors)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180212
[Lr] Data última revisão:
180212
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180113
[St] Status:MEDLINE
[do] DOI:10.1007/s11103-017-0693-6


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[PMID]:29253877
[Au] Autor:Geldreich A; Haas G; Kubina J; Bouton C; Tanguy M; Erhardt M; Keller M; Ryabova L; Dimitrova M
[Ad] Endereço:Institut de Biologie Moléculaire des Plantes, CNRS UPR2357, Université de Strasbourg, Strasbourg, France.
[Ti] Título:Formation of large viroplasms and virulence of Cauliflower mosaic virus in turnip plants depend on the N-terminal EKI sequence of viral protein TAV.
[So] Source:PLoS One;12(12):e0189062, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Cauliflower mosaic virus (CaMV) TAV protein (TransActivator/Viroplasmin) plays a pivotal role during the infection cycle since it activates translation reinitiation of viral polycistronic RNAs and suppresses RNA silencing. It is also the major component of cytoplasmic electron-dense inclusion bodies (EDIBs) called viroplasms that are particularly evident in cells infected by the virulent CaMV Cabb B-JI isolate. These EDIBs are considered as virion factories, vehicles for CaMV intracellular movement and reservoirs for CaMV transmission by aphids. In this study, focused on different TAV mutants in vivo, we demonstrate that three physically separated domains collectively participate to the formation of large EDIBs: the N-terminal EKI motif, a sequence of the MAV domain involved in translation reinitiation and a C-terminal region encompassing the zinc finger. Surprisingly, EKI mutant TAVm3, corresponding to a substitution of the EKI motif at amino acids 11-13 by three alanines (AAA), which completely abolished the formation of large viroplasms, was not lethal for CaMV but highly reduced its virulence without affecting the rate of systemic infection. Expression of TAVm3 in a viral context led to formation of small irregularly shaped inclusion bodies, mild symptoms and low levels of viral DNA and particles accumulation, despite the production of significant amounts of mature capsid proteins. Unexpectedly, for CaMV-TAVm3 the formation of viral P2-containing electron-light inclusion body (ELIB), which is essential for CaMV aphid transmission, was also altered, thus suggesting an indirect role of the EKI tripeptide in CaMV plant-to-plant propagation. This important functional contribution of the EKI motif in CaMV biology can explain the strict conservation of this motif in the TAV sequences of all CaMV isolates.
[Mh] Termos MeSH primário: Brassica napus/virologia
Caulimovirus/metabolismo
Caulimovirus/patogenicidade
Transativadores/química
Transativadores/metabolismo
[Mh] Termos MeSH secundário: Motivos de Aminoácidos
Sequência de Aminoácidos
Caulimovirus/ultraestrutura
Corpos de Inclusão Viral/metabolismo
Corpos de Inclusão Viral/ultraestrutura
Proteínas Mutantes/metabolismo
Fenótipo
Domínios Proteicos
Protoplastos/metabolismo
Transcrição Reversa/genética
Relação Estrutura-Atividade
Virulência
Replicação Viral
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Mutant Proteins); 0 (Trans-Activators); 0 (gene VI protein, Cauliflower mosaic virus)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180108
[Lr] Data última revisão:
180108
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171219
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0189062


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[PMID]:28812199
[Au] Autor:Lim S; Baek D; Igori D; Moon JS
[Ad] Endereço:Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 34141, Republic of Korea.
[Ti] Título:Complete genome sequence of a putative new caulimovirus which exists as endogenous pararetroviral sequences in Angelica dahurica.
[So] Source:Arch Virol;162(12):3837-3842, 2017 Dec.
[Is] ISSN:1432-8798
[Cp] País de publicação:Austria
[La] Idioma:eng
[Ab] Resumo:A virus isolate designated Angelica bushy stunt virus (AnBSV), provisionally representing a new species in the genus Caulimovirus, was discovered in the medicinal plant Angelica dahurica. The complete 8,300-nt genomic DNA of AnBSV had seven putative open reading frames containing conserved domains/motifs, which are typical features of caulimoviruses, and showed the greatest nucleotide sequence identity (74% identity and 27% query coverage) to a lamium leaf distortion virus isolate. Interestingly, the new caulimovirus exists as endogenous pararetroviral sequences in the host plant and is considered to have multiple defective plant genome-integrated copies that may lead to the generation of subgenomic DNA species.
[Mh] Termos MeSH primário: Angelica/virologia
Caulimovirus/genética
Caulimovirus/isolamento & purificação
Genoma Viral
Análise de Sequência de DNA
[Mh] Termos MeSH secundário: Caulimovirus/classificação
DNA Viral/química
DNA Viral/genética
Fases de Leitura Aberta
Filogenia
Homologia de Sequência
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Viral)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171113
[Lr] Data última revisão:
171113
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170817
[St] Status:MEDLINE
[do] DOI:10.1007/s00705-017-3517-8


  4 / 612 MEDLINE  
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[PMID]:28624556
[Au] Autor:Shimada A; Okumura A; Yamasaki S; Iwata Y; Koizumi N; Nishihara M; Mishiba KI
[Ad] Endereço:Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-1 Gakuen, Nakaku, Sakai, Osaka 599-8531, Japan.
[Ti] Título:A 64-bp sequence containing the GAAGA motif is essential for CaMV-35S promoter methylation in gentian.
[So] Source:Biochim Biophys Acta;1860(8):861-869, 2017 08.
[Is] ISSN:0006-3002
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:This study investigated sequence specificity and perenniality of DNA methylation in the cauliflower mosaic virus (CaMV) 35S promoter of transgenic gentian (Gentiana triflora×G. scabra) plants. Unlike conventional transgene silencing models, 35S promoter hypermethylation in gentian is species-specific and occurs irrespective of the T-DNA copy number and genomic location. Modified 35S promoters were introduced into gentian, and single-copy transgenic lines were selected for methylation analysis. Modified 35S promoter lacking a core (-90) region [35S(Δcore)] in gentian conferred hypermethylation and high levels of de novo methylation of the CpHpH/CpCpG sites in the 35S enhancer regions (-298 to -241 and -148 to -85). Therefore, promoter transcription may not be an absolute requirement for the methylation machinery. In vitro, de novo methylation persisted for more than eight years. In another modified 35S promoter, two "GAAGA" motifs (-268 to -264 and -135 to -131) were replaced by "GTTCA" in the two highly de novo methylated regions. It did not support hypermethylation and showed transgene expression. A 64-bp fragment of the 35S enhancer region (-148 to -85) was introduced into gentian and the resultant transgenic lines analyzed. The 64-bp region exhibited hypermethylation at the CpG/CpWpG sites, but the CpHpH/CpCpG methylation frequency was lower than those of the unmodified 35S- and 35S(Δcore) promoters. Nevertheless, a distinct CpHpH/CpCpG methylation peak was found in the 64-bp region of all single-copy transgenic lines. These results suggest that the 64-bp region may contain an element required for 35S methylation but insufficient for high de novo methylation compared with those in the unmodified 35S and 35S(Δcore) promoters.
[Mh] Termos MeSH primário: Caulimovirus/genética
Metilação de DNA/genética
Gentiana/genética
Regiões Promotoras Genéticas/genética
[Mh] Termos MeSH secundário: DNA Bacteriano/genética
Regulação da Expressão Gênica de Plantas/genética
Inativação Gênica/fisiologia
Plantas Geneticamente Modificadas/genética
Transcrição Genética/genética
Transgenes/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (DNA, Bacterial); 0 (T-DNA)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171026
[Lr] Data última revisão:
171026
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170619
[St] Status:MEDLINE


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[PMID]:28602750
[Au] Autor:Kadam US; Chavhan RL; Schulz B; Irudayaraj J
[Ad] Endereço:VD College of Agricultural Biotechnology (VNMKV), Latur, Maharashtra, 413512, India; Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN, 47907, USA; Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN, 47907, USA. Ele
[Ti] Título:Single molecule Raman spectroscopic assay to detect transgene from GM plants.
[So] Source:Anal Biochem;532:60-63, 2017 Sep 01.
[Is] ISSN:1096-0309
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Substantial concerns have been raised for the safety of transgenics on human health and environment. Many organizations, consumer groups, and environmental agencies advocate for stringent regulations to avoid transgene products' contamination in food cycle or in nature. Here we demonstrate a novel approach using surface enhanced Raman spectroscopy (SERS) to detect and quantify transgene from GM plants. We show a highly sensitive and accurate quantification of transgene DNA from multiple transgenic lines of Arabidopsis. The assay allows us to detect and quantify the transgenes as low as 0.10 pg without need for PCR-amplification. This technology is relatively cheap, quick, simple, and suitable for detection at low target concentration.
[Mh] Termos MeSH primário: Aminoácido Oxirredutases/genética
Arabidopsis/genética
DNA de Plantas/análise
Plantas Geneticamente Modificadas/genética
Regiões Promotoras Genéticas/genética
Análise Espectral Raman/métodos
Transgenes/fisiologia
[Mh] Termos MeSH secundário: Agrobacterium tumefaciens/enzimologia
Arabidopsis/metabolismo
Bioensaio
Caulimovirus/genética
DNA de Plantas/genética
Plantas Geneticamente Modificadas/metabolismo
Reação em Cadeia da Polimerase
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Plant); EC 1.4.- (Amino Acid Oxidoreductases); EC 1.5.1.- (nopaline synthase)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170912
[Lr] Data última revisão:
170912
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170613
[St] Status:MEDLINE


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[PMID]:28190195
[Au] Autor:Dickison V; MacKenzie TDB; Singh M; Lawrence J; Nie X
[Ad] Endereço:Fredericton Research and Development Centre, Agriculture and Agri-Food Canada, P.O. Box 20280, 850 Lincoln Road, Fredericton, NB, E3B 4Z7, Canada.
[Ti] Título:Strawberry vein banding virus isolates in eastern Canada are molecularly divergent from other isolates.
[So] Source:Arch Virol;162(6):1777-1781, 2017 Jun.
[Is] ISSN:1432-8798
[Cp] País de publicação:Austria
[La] Idioma:eng
[Ab] Resumo:The complete sequence of a strawberry vein banding virus (SVBV) isolate collected in Nova Scotia, Canada, and designated NS8, was determined. The 7,856-nucleotide circular double-stranded DNA genome contains seven open-reading frames (ORFs), which is consistent with other SVBV isolates and other members of the genus Caulimovirus. Comparison of NS8 with other whole-genome sequences retrieved from databases revealed that NS8 shares the highest sequence similarity (96.5% identity) with isolate China (accession number HE681085) and the lowest (88.3% identity) with clone pSVBV-E3 (accession number X97304). Despite the overall high sequence similarity between NS8 and China, the coat protein encoding ORF IV of NS8 shares only 90.9% sequence identity with the China isolate. Phylogenetic analysis at the complete-genome level placed NS8 and all Chinese isolates in one clade and clone pSVBV-E3 in a separate clade. Interestingly, phylogenetic analysis of all available ORF IV sequences, including those retrieved from databases and newly sequenced samples in this study from Canada, revealed three distinct clades. All Canadian isolates grouped together as one clade, pSVBV-E3 and several others from Europe, Egypt and the USA grouped as a second clade, and isolates from China formed a third clade. These results demonstrate that SVBV is more divergent than previously reported.
[Mh] Termos MeSH primário: Caulimovirus/isolamento & purificação
Fragaria/virologia
Doenças das Plantas/virologia
[Mh] Termos MeSH secundário: Sequência de Bases
Canadá
Caulimovirus/classificação
Caulimovirus/genética
China
Evolução Molecular
Genoma Viral
Dados de Sequência Molecular
Fases de Leitura Aberta
Filogenia
RNA Viral/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Viral)
[Em] Mês de entrada:1706
[Cu] Atualização por classe:170630
[Lr] Data última revisão:
170630
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170213
[St] Status:MEDLINE
[do] DOI:10.1007/s00705-017-3252-1


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[PMID]:28182975
[Au] Autor:Li Y; Sun L; Qian J; Long L; Li H; Liu Q; Cai J; Wang K
[Ad] Endereço:School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, PR China.
[Ti] Título:Fluorescent "on-off-on" switching sensor based on CdTe quantum dots coupled with multiwalled carbon nanotubes@graphene oxide nanoribbons for simultaneous monitoring of dual foreign DNAs in transgenic soybean.
[So] Source:Biosens Bioelectron;92:26-32, 2017 Jun 15.
[Is] ISSN:1873-4235
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:With the increasing concern of potential health and environmental risk, it is essential to develop reliable methods for transgenic soybean detection. Herein, a simple, sensitive and selective assay was constructed based on homogeneous fluorescence resonance energy transfer (FRET) between CdTe quantum dots (QDs) and multiwalled carbon nanotubes@graphene oxide nanoribbons (MWCNTs@GONRs) to form the fluorescent "on-off-on" switching for simultaneous monitoring dual target DNAs of promoter cauliflower mosaic virus 35s (P35s) and terminator nopaline synthase (TNOS) from transgenic soybean. The capture DNAs were immobilized with corresponding QDs to obtain strong fluorescent signals (turning on). The strong π-π stacking interaction between single-stranded DNA (ssDNA) probes and MWCNTs@GONRs led to minimal background fluorescence due to the FRET process (turning off). The targets of P35s and TNOS were recognized by dual fluorescent probes to form double-stranded DNA (dsDNA) through the specific hybridization between target DNAs and ssDNA probes. And the dsDNA were released from the surface of MWCNTs@GONRs, which leaded the dual fluorescent probes to generate the strong fluorescent emissions (turning on). Therefore, this proposed homogeneous assay can be achieved to detect P35s and TNOS simultaneously by monitoring the relevant fluorescent emissions. Moreover, this assay can distinguish complementary and mismatched nucleic acid sequences with high sensitivity. The constructed approach has the potential to be a tool for daily detection of genetically modified organism with the merits of feasibility and reliability.
[Mh] Termos MeSH primário: Compostos de Cádmio/química
Transferência Ressonante de Energia de Fluorescência/métodos
Grafite/química
Nanotubos de Carbono/química
Plantas Geneticamente Modificadas/genética
Pontos Quânticos/química
Feijão de Soja/genética
Telúrio/química
[Mh] Termos MeSH secundário: Aminoácido Oxirredutases/genética
Técnicas Biossensoriais/métodos
Caulimovirus/genética
DNA de Plantas/análise
DNA de Plantas/genética
DNA Viral/análise
DNA Viral/genética
Ácidos Nucleicos Imobilizados/química
Ácidos Nucleicos Imobilizados/genética
Nanotubos de Carbono/ultraestrutura
Óxidos/química
Proteínas de Plantas/genética
Plantas Geneticamente Modificadas/virologia
Pontos Quânticos/ultraestrutura
Reprodutibilidade dos Testes
Feijão de Soja/virologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cadmium Compounds); 0 (DNA, Plant); 0 (DNA, Viral); 0 (Immobilized Nucleic Acids); 0 (Nanotubes, Carbon); 0 (Oxides); 0 (Plant Proteins); 7782-42-5 (Graphite); EC 1.4.- (Amino Acid Oxidoreductases); EC 1.5.1.- (nopaline synthase); NQA0O090ZJ (Tellurium); STG188WO13 (cadmium telluride)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170328
[Lr] Data última revisão:
170328
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170210
[St] Status:MEDLINE


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[PMID]:28160062
[Au] Autor:Beringer J; Chen W; Garton R; Sardesai N; Wang PH; Zhou N; Gupta M; Wu H
[Ad] Endereço:Dow AgroSciences, LLC, 9330 Zionsville Road, Indianapolis, IN, 46268, USA.
[Ti] Título:Comparison of the impact of viral and plant-derived promoters regulating selectable marker gene on maize transformation and transgene expression.
[So] Source:Plant Cell Rep;36(4):519-528, 2017 Apr.
[Is] ISSN:1432-203X
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:KEY MESSAGE: The choice of promoter regulating the selectable marker gene impacts transformation efficiency, copy number and the expression of selectable marker and flanking genes in maize. Viral or plant-derived constitutive promoters are often used to regulate selectable marker genes. We compared two viral promoters, cauliflower mosaic virus (CaMV 35T) and sugarcane bacilliform virus (SCBV) with two plant promoters, rice actin1 (OsAct1) and maize ubiquitin 1 (ZmUbi1) to drive aryloxyalkanoate dioxygenase (aad-1) selectable marker gene in maize inbred line B104. ZmUbi1- and OsAct1-containing constructs demonstrated higher transformation frequencies (43.8 and 41.4%, respectively) than the two viral promoter constructs, CaMV 35T (25%) and SCBV (8%). Interestingly, a higher percentage of single copy events were recovered for SCBV (82.1%) and CaMV 35T (59.3%) promoter constructs, compared to the two plant-derived promoters, OsAct1 (40.0%), and ZmUbi1 (27.6%). Analysis of protein expression suggested that the viral promoter CaMV 35T expressed significantly higher AAD-1 protein (174.6 ng/cm ) than the OsAct1 promoter (12.6 ng/cm ) in T leaf tissue. When measured in T callus tissue, the two viral promoters both had higher expression and more variability than the two plant-derived promoters. A potential explanation for why viral promoters produce lower transformation efficiencies but higher percentages of low copy number events is discussed. In addition, viral promoters regulating aad-1 were found to influence the expression of upstream flanking genes in both T leaf and T callus tissue.
[Mh] Termos MeSH primário: Regulação da Expressão Gênica de Plantas/genética
Plantas Geneticamente Modificadas/genética
Regiões Promotoras Genéticas/genética
Transformação Genética/genética
Transgenes/genética
Zea mays/genética
[Mh] Termos MeSH secundário: Caulimovirus/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1706
[Cu] Atualização por classe:170601
[Lr] Data última revisão:
170601
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170205
[St] Status:MEDLINE
[do] DOI:10.1007/s00299-017-2099-y


  9 / 612 MEDLINE  
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[PMID]:28088748
[Au] Autor:Lv J; Miao Y; Yang J; Qin J; Li D; Yan G
[Ad] Endereço:Shanxi Normal University, Linfen 041004, PR China.
[Ti] Título:A DNA probe based on phosphorescent resonance energy transfer for detection of transgenic 35S promoter DNA.
[So] Source:Biosens Bioelectron;91:560-565, 2017 May 15.
[Is] ISSN:1873-4235
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:A QDs-DNA nano-probe was made by combining Mn-doped ZnS room-temperature phosphorescence (RTP) quantum dots (QDs) and DNA. Then an RTP sensor for quantitative detection of genetically-modified mark sequence cauliflower mosaic virus 35S promoter (Ca MV 35S) DNA was built on basis of phosphorescent resonance energy transfer (PRET). The underlying principles were that a QDs-DNA water-soluble nano-probe was built by connecting single-strand DNA to the surfaces of QDs via a ligand exchange method. This probe had good RTP performance and could well identify Ca MV 35S. Thereby, the simple, rapid and efficient detection of genetically-modified organisms was realized. With the increase of target DNA sequence, the phosphorescent intensity of QDs was gradually reduced due to the energy transfer between QDs and the organic quencher BHQ . This sensor had a detection limit of 4.03nM and a detection range of 12-300nM. Moreover, this sensor had high selectivity. This sensor could effectively detect the target DNA compared with mismatched and random sequences. Thus, this method is very promising for biological analysis.
[Mh] Termos MeSH primário: Caulimovirus/genética
Sondas de DNA/química
DNA de Cadeia Simples/química
DNA Viral/análise
Medições Luminescentes/métodos
Pontos Quânticos/química
[Mh] Termos MeSH secundário: Técnicas Biossensoriais/métodos
Sondas de DNA/genética
DNA de Cadeia Simples/genética
DNA Viral/genética
Transferência de Energia
Regiões Promotoras Genéticas
Pontos Quânticos/ultraestrutura
Sulfetos/química
Transgenes
Compostos de Zinco/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Probes); 0 (DNA, Single-Stranded); 0 (DNA, Viral); 0 (Sulfides); 0 (Zinc Compounds); KPS085631O (zinc sulfide)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170531
[Lr] Data última revisão:
170531
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170116
[St] Status:MEDLINE


  10 / 612 MEDLINE  
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[PMID]:28054967
[Au] Autor:Karuppanan K; Duhra-Gill S; Kailemia MJ; Phu ML; Lebrilla CB; Dandekar AM; Rodriguez RL; Nandi S; McDonald KA
[Ad] Endereço:Department of Chemical Engineering, University of California, Davis, CA 95616, USA. kkaruppanan@ucdavis.edu.
[Ti] Título:Expression, Purification, and Biophysical Characterization of a Secreted Anthrax Decoy Fusion Protein in Nicotiana benthamiana.
[So] Source:Int J Mol Sci;18(1), 2017 Jan 04.
[Is] ISSN:1422-0067
[Cp] País de publicação:Switzerland
[La] Idioma:eng
[Ab] Resumo:Anthrax toxin receptor-mediated drug development for blocking anthrax toxin action has received much attention in recent decades. In this study, we produced a secreted anthrax decoy fusion protein comprised of a portion of the human capillary morphogenesis gene-2 ( ) protein fused via a linker to the fragment crystallizable (Fc) domain of human immunoglobulin G1 in plants using a transient expression system. Using the Cauliflower Mosaic Virus ( ) promoter and co-expression with the p19 gene silencing suppressor, we were able to achieve a high level of recombinant CMG2-Fc-Apo (rCMG2-Fc-Apo) protein accumulation. Production kinetics were observed up to eight days post-infiltration, and maximum production of 826 mg/kg fresh leaf weight was observed on day six. Protein A affinity chromatography purification of the rCMG2-Fc-Apo protein from whole leaf extract and apoplast wash fluid showed the homodimeric form under non-reducing gel electrophoresis and mass spectrometry analysis confirmed the molecular integrity of the secreted protein. The -glycosylation pattern of purified rCMG2-Fc-Apo protein was analysed; the major portion of -glycans consists of complex type structures in both protein samples. The most abundant (>50%) -glycan structure was GlcNAc2(Xy )Man3(Fuc)GlcNAc2 in rCMG2-Fc-Apo recovered from whole leaf extract and apoplast wash fluid. High mannose -glycan structures were not detected in the apoplast wash fluid preparation, which confirmed the protein secretion. Altogether, these findings demonstrate that high-level production of rCMG2-Fc-Apo can be achieved by transient production in plants with apoplast targeting.
[Mh] Termos MeSH primário: Imunoglobulina G/genética
Plantas Geneticamente Modificadas/genética
Receptores de Peptídeos/genética
Tabaco/genética
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Antraz/metabolismo
Antraz/microbiologia
Bacillus anthracis/metabolismo
Biotecnologia
Caulimovirus/genética
Clonagem Molecular
Descoberta de Drogas
Glicosilação
Seres Humanos
Imunoglobulina G/química
Imunoglobulina G/metabolismo
Regiões Promotoras Genéticas
Receptores de Peptídeos/química
Receptores de Peptídeos/metabolismo
Proteínas Recombinantes de Fusão/química
Proteínas Recombinantes de Fusão/genética
Proteínas Recombinantes de Fusão/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (ANTXR2 protein, human); 0 (Immunoglobulin G); 0 (Receptors, Peptide); 0 (Recombinant Fusion Proteins); 0 (anthrax toxin receptors)
[Em] Mês de entrada:1704
[Cu] Atualização por classe:170413
[Lr] Data última revisão:
170413
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170106
[St] Status:MEDLINE



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