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  1 / 6911 MEDLINE  
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[PMID]:29196904
[Au] Autor:Zienkiewicz M; Krupnik T; Drozak A; Wasilewska W; Golke A; Romanowska E
[Ad] Endereço:Faculty of Biology, University of Warsaw, Miecznikowa 1, 02-096, Warsaw, Poland. maximus@biol.uw.edu.pl.
[Ti] Título:Deletion of psbQ' gene in Cyanidioschyzon merolae reveals the function of extrinsic PsbQ' in PSII.
[So] Source:Plant Mol Biol;96(1-2):135-149, 2018 Jan.
[Is] ISSN:1573-5028
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:KEY MESSAGE: We have successfully produced single-cell colonies of C. merolae mutants, lacking the PsbQ' subunit in its PSII complex by application of DTA-aided mutant selection. We have investigated the physiological changes in PSII function and structure and proposed a tentative explanation of the function of PsbQ' subunit in the PSII complex. We have improved the selectivity of the Cyanidioschyzon merolae nuclear transformation method by the introduction of diphtheria toxin genes into the transformation vector as an auxiliary selectable marker. The revised method allowed us to obtained single-cell colonies of C. merolae, lacking the gene of the PsbQ' extrinsic protein. The efficiency of gene replacement was extraordinarily high, allowing for a complete deletion of the gene of interest, without undesirable illegitimate integration events. We have confirmed the absence of PsbQ' protein at genetic and protein level. We have characterized the physiology of mutant cells and isolated PSII protein complex and concluded that PsbQ' is involved in nuclear regulation of PSII activity, by influencing several parameters of PSII function. Among these: oxygen evolving activity, partial dissociation of PsbV, regulation of dimerization, downsizing of phycobilisomes rods and regulation of zeaxanthin abundance. The adaptation of cellular physiology appeared to favorite upregulation of PSII and concurrent downregulation of PSI, resulting in an imbalance of energy distribution, decrease of photosynthesis and inhibition of cell proliferation.
[Mh] Termos MeSH primário: Fotossíntese/genética
Complexo de Proteína do Fotossistema II/genética
Complexo de Proteína do Fotossistema II/metabolismo
[Mh] Termos MeSH secundário: Cloranfenicol O-Acetiltransferase/genética
Cloranfenicol O-Acetiltransferase/metabolismo
Rodófitas/genética
Rodófitas/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Photosystem II Protein Complex); EC 2.3.1.28 (Chloramphenicol O-Acetyltransferase)
[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:171203
[St] Status:MEDLINE
[do] DOI:10.1007/s11103-017-0685-6


  2 / 6911 MEDLINE  
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[PMID]:28813510
[Au] Autor:Wang R; Lin C; Lin J; Pang X; Liu X; Zhang C; Lin J; Chen L
[Ad] Endereço:State Key Laboratory of Microbial Technology, Shandong University, Jinan, China.
[Ti] Título:Construction of novel pJRD215-derived plasmids using chloramphenicol acetyltransferase (cat) gene as a selection marker for Acidithiobacillus caldus.
[So] Source:PLoS One;12(8):e0183307, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Acidithiobacillus caldus, a Gram-negative, chemolithotrophic sulfur-oxidizing bacterium, is widely applied in bioleaching. The absence of an ideal selection marker has become a major obstacle to achieve high efficiency of the gene transfer system for A. caldus. Plasmid pJRD215, widely used in Acidithiobacillus spp., has severe drawbacks in molecular manipulations and potential biosafety issues due to its mobility. Therefore, finding a new selection marker and constructing new plasmids have become an urgent and fundamental work for A. caldus. RESULTS: Effective inhibitory effect of chloramphenicol on the growth of A. caldus was elucidated for the first time. The P2-cat gene cassette, including a chloramphenicol acetyltransferase gene (cat) from plasmid pACBSR and a promoter (P2) upstream of the tetracycline resistance gene on pBR322, was designed, chloramphenicol acetyltransferase was expressed in A. caldus, and the enzyme activity was assessed. A new vector pSDU1 carrying the replication and mobilization regions derived from pJRD215, the P2-cat gene cassette and a multiple cloning site from pUC19 was successfully constructed. Compared with pJRD215, pSDU1 had a 27-fold increase in electrotransformation efficiency (30.43±0.88×104 CFU/µg DNA for pSDU1 and 1.09±0.11×104 CFU/µg DNA for pJRD215), better carrying capacity and could offer more convenience for the restriction enzyme digestion. In addition, the generated plasmid pSDU1Δmob, a novel non-mobilizable derivative of pSDU1 lacking some DNA sequences involved in the mobilization process, had increased copy number in A. caldus and lost its mobility for biosafety considerations. Both pSDU1 and pSDU1Δmob exhibited stable maintenance in A. caldus within 50 passages. However, further deletion of orfEF region involved in regulating repAC operon resulted in a negative effect on transformation efficiency, copy number and stability of plasmid pSDU1ΔmobΔorfEF in A. caldus. CONCLUSION: Chloramphenicol was proved to be an ideal selection marker for A. caldus. Novel plasmids carrying cat gene were constructed. The utilization of these vectors will undoubtedly facilitate efficient genetic manipulations and accelerate the research progress in A. caldus.
[Mh] Termos MeSH primário: Acidithiobacillus/metabolismo
Cloranfenicol O-Acetiltransferase/genética
Plasmídeos/genética
[Mh] Termos MeSH secundário: Acidithiobacillus/efeitos dos fármacos
Acidithiobacillus/genética
Antibacterianos/farmacologia
Proteínas de Bactérias/genética
Cloranfenicol/farmacologia
Vetores Genéticos/genética
Regiões Promotoras Genéticas/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anti-Bacterial Agents); 0 (Bacterial Proteins); 66974FR9Q1 (Chloramphenicol); EC 2.3.1.28 (Chloramphenicol O-Acetyltransferase)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171013
[Lr] Data última revisão:
171013
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170817
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0183307


  3 / 6911 MEDLINE  
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[PMID]:28370370
[Au] Autor:Chai M; Liu B; Sun F; Wei P; Chen P; Xu L; Luo SZ
[Ad] Endereço:Beijing Key Laboratory of Bioprocess College of Life Science and Technology, Beijing, University of Chemical Technology, Beijing, 100029, China.
[Ti] Título:Insights into the transmembrane helix associations of kit ligand by molecular dynamics simulation and TOXCAT.
[So] Source:Proteins;85(7):1362-1370, 2017 Jul.
[Is] ISSN:1097-0134
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Kit ligand (KITL) plays important roles in cell proliferation, differentiation, and survival via interaction with its receptor Kit. The previous studies demonstrated that KITL formed a noncovalent homodimer through transmembrane (TM) domain; however, the undergoing mechanism of transmembrane association that determines KITL TM dimerization is still not clear. Herein, molecular dynamics (MD) simulation strategy and TOXCAT assay were combined to characterize the dimerization interface and structure of KITL TM in details. KITL TM formed a more energetically favorable noncovalent dimer through a conserved SxxxGxxxG motif in the MD simulation. Furthermore, the TOXCAT results demonstrated that KITL TM self-associated strongly in the bilayer membrane environment. Mutating any one of the small residues Ser11, Gly15 or Gly19 to Ile disrupted KITL TM dimerization dramatically, which further validated our MD simulation results. In addition, our results showed that Tyr22 could help to stabilize the TM interactions via interacting with the phosphoric group in the bilayer membrane. Pro7 did not induce helix kinks or swivel angles in KITL TM, but it was related with the pitch of the turn around this residue so as to affect the dimer formation. Combining the results of computer modeling and experimental mutagenesis studies on the KITL TM provide new insights for the transmembrane helix association of KITL dimerization. Proteins 2017; 85:1362-1370. © 2017 Wiley Periodicals, Inc.
[Mh] Termos MeSH primário: Cloranfenicol O-Acetiltransferase/metabolismo
Simulação de Dinâmica Molecular
Proteínas Recombinantes de Fusão/química
Fator de Células-Tronco/química
[Mh] Termos MeSH secundário: 1,2-Dipalmitoilfosfatidilcolina/química
Sequência de Aminoácidos
Animais
Sítios de Ligação
Bovinos
Cloranfenicol O-Acetiltransferase/genética
Clonagem Molecular
Cães
Escherichia coli/genética
Escherichia coli/metabolismo
Expressão Gênica
Genes Reporter
Seres Humanos
Cinética
Bicamadas Lipídicas/química
Camundongos
Mutação
Ligação Proteica
Conformação Proteica em alfa-Hélice
Conformação Proteica em Folha beta
Multimerização Proteica
Ratos
Proteínas Recombinantes de Fusão/genética
Proteínas Recombinantes de Fusão/metabolismo
Proteínas Recombinantes/química
Proteínas Recombinantes/genética
Proteínas Recombinantes/metabolismo
Alinhamento de Sequência
Fator de Células-Tronco/genética
Fator de Células-Tronco/metabolismo
Especificidade por Substrato
Suínos
Termodinâmica
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Lipid Bilayers); 0 (Recombinant Fusion Proteins); 0 (Recombinant Proteins); 0 (Stem Cell Factor); 2644-64-6 (1,2-Dipalmitoylphosphatidylcholine); EC 2.3.1.28 (Chloramphenicol O-Acetyltransferase)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170728
[Lr] Data última revisão:
170728
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170404
[St] Status:MEDLINE
[do] DOI:10.1002/prot.25297


  4 / 6911 MEDLINE  
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[PMID]:28364624
[Au] Autor:Shen Z; Mustapha A; Lin M; Zheng G
[Ad] Endereço:Department of Agriculture and Environmental Sciences, Lincoln University of Missouri, 904 Chestnut Street, Jefferson City, MO 65101, USA; Food Science Program, University of Missouri, Columbia, MO 65211, USA.
[Ti] Título:Biocontrol of the internalization of Salmonella enterica and Enterohaemorrhagic Escherichia coli in mung bean sprouts with an endophytic Bacillus subtilis.
[So] Source:Int J Food Microbiol;250:37-44, 2017 Jun 05.
[Is] ISSN:1879-3460
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Internalization of Salmonella enterica and enterohaemorrhagic Escherichia coli (EHEC) in seed sprouts poses a health risk to consumers, and the conventional sanitization methods are not always effective to reduce this risk. This study initiated a biocontrol approach to limit the internalization using endophytic Bacillus subtilis strains, which were isolated from the inner tissue of mung bean seeds or lettuce stems. By using the deferred agar method, 12 strains of B. subtilis out of 94 putative Bacillus isolates displayed inhibitory activity against at least one of the pathogenic indicators, S. enterica Typhimurium ATCC 14028 and E. coli O157:H7 505B. Two B. subtilis isolates (LCA1 and M24) showed a broad inhibitory spectrum against multiple strains of S. enterica and EHEC, Staphylococcus aureus sp., Klebsiella pneumoniae ATCC 700603, and Listeria monocytogenes Scott A, while the laboratory B. subtilis strain 168 was only moderately inhibitory against L. monocytogenes. To facilitate the tracking of the three B. subtilis strains (LCA1, M24, and 168) in the mung bean sprouts, the three strains were genetically engineered to carry the chloramphenicol acetyltransferase (cat), generating the strains LCA1-cat, M24-cat, and 168-cat, respectively. Data of the study using the cat-tagged strains demonstrated that both the two vegetable-associated and the laboratory B. subtilis strains could internalize in mung bean sprouts during the sprouting, but the latter displayed about 1.2 lg CFU/g of seeds lower in internalization. Overall, the presence of the three B. subtilis strains could significantly reduce the internalization of S. enterica or EHEC cocktail in mung bean sprouts during the sprouting. Among them, LCA1 showed the greatest inhibition against the EHEC cocktails with a reduction of about 2.0lg CFU/g of seeds by the end of sprouting (day 5), while 168 had the smallest reduction at about 0.6lg CFU/g of seeds. In addition, the three strains demonstrated a similar inhibition against the S. enterica cocktails by a reduction of about 1.1-1.4lg CFU/g of seeds by day 5. Results of this study suggest that the source (native vs. alien) of B. subtilis isolates may not affect the efficacy of the inhibition, but it might be affected by the production of antimicrobial substance and/or nutrition/space competition. The results also indicate that strain LCA1 may be useful as a biocontrol agent to reduce Salmonella and EHEC contamination in seed sprouts.
[Mh] Termos MeSH primário: Bacillus subtilis/metabolismo
Agentes de Controle Biológico/metabolismo
Escherichia coli O157/crescimento & desenvolvimento
Contaminação de Alimentos/prevenção & controle
Klebsiella pneumoniae/crescimento & desenvolvimento
Listeria monocytogenes/crescimento & desenvolvimento
Salmonella typhimurium/crescimento & desenvolvimento
Staphylococcus aureus/crescimento & desenvolvimento
Vigna/microbiologia
[Mh] Termos MeSH secundário: Bacillus subtilis/genética
Cloranfenicol O-Acetiltransferase/genética
Contagem de Colônia Microbiana
Escherichia coli O157/isolamento & purificação
Microbiologia de Alimentos
Germinação
Klebsiella pneumoniae/isolamento & purificação
Listeria monocytogenes/isolamento & purificação
Salmonella typhimurium/isolamento & purificação
Sementes/microbiologia
Staphylococcus aureus/isolamento & purificação
Verduras/microbiologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Biological Control Agents); EC 2.3.1.28 (Chloramphenicol O-Acetyltransferase)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170928
[Lr] Data última revisão:
170928
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170402
[St] Status:MEDLINE


  5 / 6911 MEDLINE  
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[PMID]:28270274
[Au] Autor:Medina-Aparicio L; Rebollar-Flores JE; Beltrán-Luviano AA; Vázquez A; Gutiérrez-Ríos RM; Olvera L; Calva E; Hernández-Lucas I
[Ad] Endereço:Departamento de Microbiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Cuernavaca, Morelos 62210, Mexico.
[Ti] Título:CRISPR-Cas system presents multiple transcriptional units including antisense RNAs that are expressed in minimal medium and upregulated by pH in Salmonella enterica serovar Typhi.
[So] Source:Microbiology;163(2):253-265, 2017 Feb.
[Is] ISSN:1465-2080
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The CRISPR-Cas system is involved in bacterial immunity, virulence, gene regulation, biofilm formation and sporulation. In Salmonella enterica serovar Typhi, this system consists of five transcriptional units including antisense RNAs. It was determined that these genetic elements are expressed in minimal medium and are up-regulated by pH. In addition, a transcriptional characterization of cas3 and ascse2-1 is included herein.
[Mh] Termos MeSH primário: Proteínas Associadas a CRISPR/genética
Sistemas CRISPR-Cas/genética
DNA Helicases/genética
Regulação Bacteriana da Expressão Gênica/genética
RNA Antissenso/genética
Salmonella typhi/genética
[Mh] Termos MeSH secundário: Cloranfenicol O-Acetiltransferase/metabolismo
Ensaio de Desvio de Mobilidade Eletroforética
Transcrição Genética/genética
Ativação Transcricional/genética
Regulação para Cima/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (CRISPR-Associated Proteins); 0 (RNA, Antisense); EC 2.3.1.28 (Chloramphenicol O-Acetyltransferase); EC 3.6.4.- (DNA Helicases)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170926
[Lr] Data última revisão:
170926
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170309
[St] Status:MEDLINE
[do] DOI:10.1099/mic.0.000414


  6 / 6911 MEDLINE  
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[PMID]:28199699
[Au] Autor:Verner-Jeffreys DW; Brazier T; Perez RY; Ryder D; Card RM; Welch TJ; Hoare R; Ngo T; McLaren N; Ellis R; Bartie KL; Feist SW; Rowe WMP; Adams A; Thompson KD
[Ad] Endereço:Cefas Weymouth laboratory, The Nothe, Barrack Road, Weymouth DT4 8UB, UK.
[Ti] Título:Detection of the florfenicol resistance gene floR in Chryseobacterium isolates from rainbow trout. Exception to the general rule?
[So] Source:FEMS Microbiol Ecol;93(4), 2017 Apr 01.
[Is] ISSN:1574-6941
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Bacteria from the family Flavobacteriaceae often show low susceptibility to antibiotics. With the exception of two Chryseobacterium spp. isolates that were positive for the florfenicol resistance gene floR, no clinical resistance genes were identified by microarray in 36 Flavobacteriaceae isolates from salmonid fish that could grow in ≥ 4 mg/L florfenicol. Whole genome sequence analysis of the floR positive isolates revealed the presence of a region that contained the antimicrobial resistance genes floR, a tet(X) tetracycline resistance gene, a streptothricin resistance gene and a chloramphenicol acetyltransferase gene. In silico analysis of 377 published genomes for Flavobacteriaceae isolates from a range of sources confirmed that well-characterised resistance gene cassettes were not widely distributed in bacteria from this group. Efflux pump-mediated decreased susceptibility to a range of antimicrobials was confirmed in both floR positive isolates using an efflux pump inhibitor (phenylalanine-arginine ß-naphthylamide) assay. The floR isolates possessed putative virulence factors, including production of siderophores and haemolysins, and were mildly pathogenic in rainbow trout. Results support the suggestion that, despite the detection of floR, susceptibility to antimicrobials in Flavobacteriaceae is mostly mediated via intrinsic mechanisms rather than the horizontally acquired resistance genes more normally associated with Gram-negative bacterial pathogens such as Enterobacteriaceae.
[Mh] Termos MeSH primário: Antibacterianos/farmacologia
Proteínas de Bactérias/genética
Chryseobacterium/efeitos dos fármacos
Chryseobacterium/genética
Oncorhynchus mykiss/microbiologia
Tianfenicol/análogos & derivados
[Mh] Termos MeSH secundário: Acetiltransferases/genética
Animais
Proteínas de Transporte/antagonistas & inibidores
Proteínas de Transporte/genética
Cloranfenicol O-Acetiltransferase/genética
Chryseobacterium/isolamento & purificação
Genoma Bacteriano/genética
Proteínas Hemolisinas/biossíntese
Seres Humanos
Testes de Sensibilidade Microbiana
Fenilalanina/análogos & derivados
Fenilalanina/farmacologia
Reação em Cadeia da Polimerase
Sideróforos/biossíntese
Resistência a Tetraciclina/genética
Tianfenicol/farmacologia
Fatores de Virulência/biossíntese
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anti-Bacterial Agents); 0 (Bacterial Proteins); 0 (Carrier Proteins); 0 (Hemolysin Proteins); 0 (Siderophores); 0 (Virulence Factors); 47E5O17Y3R (Phenylalanine); 740-57-8 (phenylalanine-beta-naphthylamide); 9J97307Y1H (florfenicol); EC 2.3.1.- (Acetyltransferases); EC 2.3.1.- (streptothricin acetyltransferase); EC 2.3.1.28 (Chloramphenicol O-Acetyltransferase); FLQ7571NPM (Thiamphenicol)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171003
[Lr] Data última revisão:
171003
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170216
[St] Status:MEDLINE
[do] DOI:10.1093/femsec/fix015


  7 / 6911 MEDLINE  
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[PMID]:27796719
[Au] Autor:Zienkiewicz M; Krupnik T; Drozak A; Golke A; Romanowska E
[Ad] Endereço:Department of Molecular Plant Physiology, Faculty of Biology, University of Warsaw, ul. Miecznikowa 1, 02-096, Warsaw, Poland. maximus@biol.uw.edu.pl.
[Ti] Título:Transformation of the Cyanidioschyzon merolae chloroplast genome: prospects for understanding chloroplast function in extreme environments.
[So] Source:Plant Mol Biol;93(1-2):171-183, 2017 Jan.
[Is] ISSN:1573-5028
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:KEY MESSAGE: We have successfully transformed an exthemophilic red alga with the chloramphenicol acetyltransferase gene, rendering this organism insensitive to its toxicity. Our work paves the way to further work with this new modelorganism. Here we report the first successful attempt to achieve a stable, under selectable pressure, chloroplast transformation in Cyanidioschizon merolae-an extremophilic red alga of increasing importance as a new model organism. The following protocol takes advantage of a double homologous recombination phenomenon in the chloroplast, allowing to introduce an exogenous, selectable gene. For that purpose, we decided to use chloramphenicol acetyltransferase (CAT), as chloroplasts are particularly vulnerable to chloramphenicol lethal effects (Zienkiewicz et al. in Protoplasma, 2015, doi: 10.1007/s00709-015-0936-9 ). We adjusted two methods of DNA delivery: the PEG-mediated delivery and the biolistic bombardment based delivery, either of these methods work sufficiently with noticeable preference to the former. Application of a codon-optimized sequence of the cat gene and a single colony selection yielded C. merolae strains, capable of resisting up to 400 µg/mL of chloramphenicol. Our method opens new possibilities in production of site-directed mutants, recombinant proteins and exogenous protein overexpression in C. merolae-a new model organism.
[Mh] Termos MeSH primário: Cloroplastos/genética
Genoma de Cloroplastos
Rodófitas/genética
[Mh] Termos MeSH secundário: Biolística
Cloranfenicol O-Acetiltransferase/genética
Cloroplastos/fisiologia
Recombinação Homóloga
Plantas Geneticamente Modificadas
Transformação Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
EC 2.3.1.28 (Chloramphenicol O-Acetyltransferase)
[Em] Mês de entrada:1702
[Cu] Atualização por classe:170224
[Lr] Data última revisão:
170224
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161101
[St] Status:MEDLINE
[do] DOI:10.1007/s11103-016-0554-8


  8 / 6911 MEDLINE  
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[PMID]:26715590
[Au] Autor:Zienkiewicz M; Krupnik T; Drozak A; Golke A; Romanowska E
[Ad] Endereço:Warsaw University, Warsaw, Poland. maximus@biol.uw.edu.pl.
[Ti] Título:Chloramphenicol acetyltransferase-a new selectable marker in stable nuclear transformation of the red alga Cyanidioschyzon merolae.
[So] Source:Protoplasma;254(1):587-596, 2017 Jan.
[Is] ISSN:1615-6102
[Cp] País de publicação:Austria
[La] Idioma:eng
[Ab] Resumo:In this study, we have shown the applicability of chloramphenicol acetyltransferase as a new and convenient selectable marker for stable nuclear transformation as well as potential chloroplast transformation of Cyanidioschyzon merolae-a new model organism, which offers unique opportunities for studding the mitochondrial and plastid physiology as well as various evolutionary, structural, and functional features of the photosynthetic apparatus.
[Mh] Termos MeSH primário: Núcleo Celular/metabolismo
Cloranfenicol O-Acetiltransferase/metabolismo
Resistência ao Cloranfenicol/genética
Rodófitas/genética
Transformação Genética
[Mh] Termos MeSH secundário: Marcadores Genéticos
Mutação/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Genetic Markers); EC 2.3.1.28 (Chloramphenicol O-Acetyltransferase)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:171012
[Lr] Data última revisão:
171012
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:151231
[St] Status:MEDLINE
[do] DOI:10.1007/s00709-015-0936-9


  9 / 6911 MEDLINE  
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[PMID]:27592072
[Au] Autor:Li L; Olsen RH; Shi L; Ye L; He J; Meng H
[Ad] Endereço:School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, Guangdong, PR China; Department of Veterinary Disease Biology, Faculty Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.
[Ti] Título:Characterization of a plasmid carrying cat, ermB and tetS genes in a foodborne Listeria monocytogenes strain and uptake of the plasmid by cariogenic Streptococcus mutans.
[So] Source:Int J Food Microbiol;238:68-71, 2016 Dec 05.
[Is] ISSN:1879-3460
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:A multi-drug resistant (MDR) Listeria monocytogenes isolate (serotype 1/2c) was recovered from a quick-frozen rice flour product collected from Langfang city in northern China. PCR screening identified the presence of cat, ermB and tetS genes. The plasmid profile of the strain showed the presence of an approximately 22.4-kb plasmid. Curing of this plasmid resulted in the loss of cat, ermB and tetS genes and increased susceptibility to several antibiotics, suggesting the involvement of the plasmid in multiple antibiotic resistances. Moreover, the plasmid was able to be uptaken by human oral pathogen Streptococcus mutans by natural transformation and resulted in the acquiring of multiple resistances in the transconjugants. This study contributes to our knowledge on acquired multi-drug resistance in foodborne pathogenic L.monocytogenes, which will add to a better understanding of effective clinical management of listeriosis.
[Mh] Termos MeSH primário: Resistência Microbiana a Medicamentos/genética
Listeria monocytogenes/genética
Plasmídeos/genética
Streptococcus mutans/genética
[Mh] Termos MeSH secundário: Antibacterianos/farmacologia
Proteínas de Bactérias/genética
China
Cloranfenicol O-Acetiltransferase/genética
Resistência a Múltiplos Medicamentos/genética
Listeria monocytogenes/efeitos dos fármacos
Proteínas de Membrana Transportadoras/genética
Metiltransferases/genética
Testes de Sensibilidade Microbiana
Streptococcus mutans/efeitos dos fármacos
Transformação Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anti-Bacterial Agents); 0 (Bacterial Proteins); 0 (Membrane Transport Proteins); 0 (Tet(S) protein, Listeria monocytogenes); EC 2.1.1.- (Methyltransferases); EC 2.3.1.28 (Chloramphenicol O-Acetyltransferase)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170817
[Lr] Data última revisão:
170817
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160905
[St] Status:MEDLINE


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[PMID]:27453198
[Au] Autor:Armstrong CR; Senes A
[Ad] Endereço:Dept of Biochemistry, University of Wisconsin-Madison, 433 Babcock Dr., Madison, WI 53706, United States.
[Ti] Título:Screening for transmembrane association in divisome proteins using TOXGREEN, a high-throughput variant of the TOXCAT assay.
[So] Source:Biochim Biophys Acta;1858(11):2573-2583, 2016 11.
[Is] ISSN:0006-3002
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:TOXCAT is a widely used genetic assay to study interactions of transmembrane helices within the inner membrane of the bacterium Escherichia coli. TOXCAT is based on a fusion construct that links a transmembrane domain of interest with a cytoplasmic DNA-binding domain from the Vibrio cholerae ToxR protein. Interaction driven by the transmembrane domain results in dimerization of the ToxR domain, which, in turn, activates the expression of the reporter gene chloramphenicol acetyl transferase (CAT). Quantification of CAT is used as a measure of the ability of the transmembrane domain to self-associate. Because the quantification of CAT is relatively laborious, we developed a high-throughput variant of the assay, TOXGREEN, based on the expression of super-folded GFP and detection of fluorescence directly in unprocessed cell cultures. Careful side-by-side comparison of TOXCAT and TOXGREEN demonstrates that the methods have comparable response, dynamic range, sensitivity and intrinsic variability both in LB and minimal media. The greatly enhanced workflow makes TOXGREEN much more scalable and ideal for screening, since hundreds of constructs can be rapidly assessed in 96 well plates. Even for small scale investigations, TOXGREEN significantly reduces time, labor and cost associated with the procedure. We demonstrate applicability with a large screening for self-association among the transmembrane domains of bitopic proteins of the divisome (FtsL, FtsB, FtsQ, FtsI, FtsN, ZipA and EzrA) belonging to 11 bacterial species. The analysis confirms a previously reported tendency for FtsB to self-associate, and suggests that the transmembrane domains of ZipA, EzrA and FtsN may also possibly oligomerize.
[Mh] Termos MeSH primário: Proteínas de Bactérias/genética
Proteínas de Ciclo Celular/genética
Proteínas de Ligação a DNA/genética
Proteínas de Escherichia coli/genética
Escherichia coli/genética
Proteínas de Fluorescência Verde/genética
Ensaios de Triagem em Larga Escala
Fatores de Transcrição/genética
[Mh] Termos MeSH secundário: Proteínas de Bactérias/metabolismo
Proteínas de Ciclo Celular/metabolismo
Cloranfenicol O-Acetiltransferase/genética
Cloranfenicol O-Acetiltransferase/metabolismo
Proteínas de Ligação a DNA/metabolismo
Escherichia coli/classificação
Escherichia coli/metabolismo
Proteínas de Escherichia coli/metabolismo
Expressão Gênica
Genes Reporter
Proteínas de Fluorescência Verde/metabolismo
Proteínas de Membrana/genética
Proteínas de Membrana/metabolismo
Filogenia
Ligação Proteica
Dobramento de Proteína
Domínios e Motivos de Interação entre Proteínas
Isoformas de Proteínas/genética
Isoformas de Proteínas/metabolismo
Multimerização Proteica
Estrutura Secundária de Proteína
Proteínas Recombinantes/genética
Proteínas Recombinantes/metabolismo
Sensibilidade e Especificidade
Fatores de Transcrição/metabolismo
Vibrio cholerae/classificação
Vibrio cholerae/genética
Vibrio cholerae/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 (Bacterial Proteins); 0 (Cell Cycle Proteins); 0 (DNA-Binding Proteins); 0 (Escherichia coli Proteins); 0 (FtsB protein, E coli); 0 (Membrane Proteins); 0 (Protein Isoforms); 0 (Recombinant Proteins); 0 (Transcription Factors); 0 (toxR protein, Vibrio cholerae); 147336-22-9 (Green Fluorescent Proteins); EC 2.3.1.28 (Chloramphenicol O-Acetyltransferase)
[Em] Mês de entrada:1706
[Cu] Atualização por classe:171101
[Lr] Data última revisão:
171101
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160726
[St] Status:MEDLINE



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