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[PMID]:28352998
[Au] Autor:Xie Z; Zhang Z; Cao Z; Chen M; Li P; Liu W; Qin H; Zhao X; Tao Y; Chen Y
[Ad] Endereço:State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, People's Republic of China.
[Ti] Título:An external substrate-free blue/white screening system in Escherichia coli.
[So] Source:Appl Microbiol Biotechnol;101(9):3811-3820, 2017 May.
[Is] ISSN:1432-0614
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:Since the lacZα-based blue/white screening system was introduced to molecular biology, several different visual reporter systems were developed and used for various purposes in Escherichia coli. A common limit to the existent visual reporter systems is that an extracellular chromogenic substrate has to be added for the visible pigment production. In this study, we developed a new blue/white screening system based on a non-ribosomal peptide synthetase encoded by idgS from Streptomyces and a phosphopantetheinyl transferase encoded by sfp from Bacillus. When IdgS is activated from an apo-form to a holo-form via a posttranslational modification catalyzed by Sfp, it can synthesize a blue pigment indigoidine using L-glutamine, the amino acid abundant in cells, as a substrate. The new blue/white screening system contains a recipient E. coli strain with an optimized idgS gene cassette and a cloning vector harboring an sfp gene with an in-frame insertion of a multiple cloning site close to its N-terminal. We demonstrated that the IdgS/Sfp-based blue/white screening system is a powerful alternative to the lacZα-based screening system, which does not require any external substrate addition.
[Mh] Termos MeSH primário: Clonagem Molecular
Escherichia coli/genética
Testes Genéticos/métodos
Vetores Genéticos
Genética Microbiana/métodos
Biologia Molecular/métodos
Pigmentos Biológicos/análise
[Mh] Termos MeSH secundário: Bacillus/enzimologia
Bacillus/genética
Proteínas de Bactérias/genética
Proteínas de Bactérias/metabolismo
Cor
Peptídeo Sintases/genética
Peptídeo Sintases/metabolismo
Streptomyces/enzimologia
Streptomyces/genética
Transferases (Outros Grupos de Fosfato Substituídos)/genética
Transferases (Outros Grupos de Fosfato Substituídos)/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Bacterial Proteins); 0 (Pigments, Biological); 0 (phosphopantetheinyl transferase); EC 2.7.8.- (Transferases (Other Substituted Phosphate Groups)); EC 6.3.2.- (Peptide Synthases); EC 6.3.2.- (non-ribosomal peptide synthase)
[Em] Mês de entrada:1704
[Cu] Atualização por classe:170428
[Lr] Data última revisão:
170428
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170330
[St] Status:MEDLINE
[do] DOI:10.1007/s00253-017-8252-2


  2 / 7593 MEDLINE  
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[PMID]:28348028
[Au] Autor:Weaver KE; Chen Y; Miiller EM; Johnson JN; Dangler AA; Manias DA; Clem AM; Schjodt DJ; Dunny GM
[Ad] Endereço:Division of Basic Biomedical Sciences, Sanford School of Medicine, University of South Dakota, Vermillion, South Dakota, USA kweaver@usd.edu.
[Ti] Título:Examination of Enterococcus faecalis Toxin-Antitoxin System Toxin Fst Function Utilizing a Pheromone-Inducible Expression Vector with Tight Repression and Broad Dynamic Range.
[So] Source:J Bacteriol;199(12), 2017 Jun 15.
[Is] ISSN:1098-5530
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Tools for regulated gene expression in are extremely limited. In this report, we describe the construction of an expression vector for , designated pCIE, utilizing the P pheromone-responsive promoter of plasmid pCF10. We demonstrate that this promoter is tightly repressed, responds to nanogram quantities of the peptide pheromone, and has a large dynamic range. To demonstrate its utility, the promoter was used to control expression of the toxic peptides of two family toxin-antitoxin (TA) loci present in , of the pAD1 plasmid and located on the chromosome. The results demonstrated differences in the modes of regulation of toxin expression and in the effects of toxins of these two related systems. We anticipate that this vector will be useful for further investigation of TA system function as well as the regulated expression of other genes in is an important nosocomial pathogen and a model organism for examination of the genetics and physiology of Gram-positive cocci. While numerous genetic tools have been generated for the manipulation of this organism, vectors for the regulated expression of cloned genes remain limited by high background expression and the use of inducers with undesirable effects on the cell. Here we demonstrate that the P pheromone-responsive promoter is repressed tightly enough to allow cloning of TA system toxins and evaluate their effects at very low induction levels. This tool will allow us to more fully examine TA system function in and to further elucidate its potential roles in cell physiology.
[Mh] Termos MeSH primário: Toxinas Bacterianas/genética
Toxinas Bacterianas/metabolismo
Enterococcus faecalis/genética
Enterococcus faecalis/metabolismo
Regulação da Expressão Gênica/efeitos dos fármacos
Vetores Genéticos
Feromônios/metabolismo
[Mh] Termos MeSH secundário: Genética Microbiana/métodos
Biologia Molecular/métodos
Regiões Promotoras Genéticas
Proteínas Recombinantes/genética
Proteínas Recombinantes/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Bacterial Toxins); 0 (Pheromones); 0 (Recombinant Proteins); 0 (fst toxin, Enterococcus faecalis)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:171126
[Lr] Data última revisão:
171126
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170329
[St] Status:MEDLINE


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[PMID]:28320886
[Au] Autor:Ryu MH; Fomicheva A; Moskvin OV; Gomelsky M
[Ad] Endereço:Department of Molecular Biology, University of Wyoming, Laramie, Wyoming, USA.
[Ti] Título:Optogenetic Module for Dichromatic Control of c-di-GMP Signaling.
[So] Source:J Bacteriol;199(18), 2017 Sep 15.
[Is] ISSN:1098-5530
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Many aspects of bacterial physiology and behavior, including motility, surface attachment, and the cell cycle, are controlled by cyclic di-GMP (c-di-GMP)-dependent signaling pathways on the scale of seconds to minutes. Interrogation of such processes in real time requires tools for introducing rapid and reversible changes in intracellular c-di-GMP levels. Inducing the expression of genes encoding c-di-GMP-synthetic (diguanylate cyclases) and -degrading (c-di-GMP phosphodiesterase) enzymes by chemicals may not provide adequate temporal control. In contrast, light-controlled diguanylate cyclases and phosphodiesterases can be quickly activated and inactivated. A red/near-infrared-light-regulated diguanylate cyclase, BphS, was engineered previously, yet a complementary light-activated c-di-GMP phosphodiesterase has been lacking. In search of such a phosphodiesterase, we investigated two homologous proteins from and , designated BldP, which contain C-terminal EAL-BLUF modules, where EAL is a c-di-GMP phosphodiesterase domain and BLUF is a blue light sensory domain. Characterization of the BldP proteins in and showed that they possess light-activated c-di-GMP phosphodiesterase activities. Interestingly, light activation in both enzymes was dependent on oxygen levels. The truncated EAL-BLUF fragment from BldP lacked phosphodiesterase activity, whereas a similar fragment from BldP, designated EB1, possessed such activity that was highly (>30-fold) upregulated by light. Following light withdrawal, EB1 reverted to the inactive ground state with a half-life of ∼6 min. Therefore, the blue-light-activated phosphodiesterase EB1 can be used in combination with the red/near-infrared-light-regulated diguanylate cyclase BphS for the bidirectional regulation of c-di-GMP-dependent processes in as well as other bacterial and nonbacterial cells. Regulation of motility, attachment to surfaces, the cell cycle, and other bacterial processes controlled by the c-di-GMP signaling pathways occur at a fast (seconds-to-minutes) pace. Interrogation of these processes at high temporal and spatial resolution using chemicals is difficult or impossible, while optogenetic approaches may prove useful. We identified and characterized a robust, blue-light-activated c-di-GMP phosphodiesterase (hydrolase) that complements a previously engineered red/near-infrared-light-regulated diguanylate cyclase (c-di-GMP synthase). These two enzymes form a dichromatic module for manipulating intracellular c-di-GMP levels in bacterial and nonbacterial cells.
[Mh] Termos MeSH primário: GMP Cíclico/análogos & derivados
Escherichia coli/metabolismo
Escherichia coli/efeitos da radiação
Genética Microbiana/métodos
Optogenética/métodos
Diester Fosfórico Hidrolases/metabolismo
Transdução de Sinais
[Mh] Termos MeSH secundário: Alphaproteobacteria/enzimologia
Alphaproteobacteria/genética
Chromatiaceae/enzimologia
Chromatiaceae/genética
GMP Cíclico/metabolismo
Escherichia coli/enzimologia
Escherichia coli/genética
Luz
Diester Fosfórico Hidrolases/genética
Proteínas Recombinantes/genética
Proteínas Recombinantes/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Recombinant Proteins); 61093-23-0 (bis(3',5')-cyclic diguanylic acid); EC 3.1.4.- (Phosphoric Diester Hydrolases); H2D2X058MU (Cyclic GMP)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170907
[Lr] Data última revisão:
170907
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170322
[St] Status:MEDLINE


  4 / 7593 MEDLINE  
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[PMID]:28258143
[Au] Autor:Liu M; Zhang L; Huang L; Biville F; Zhu D; Wang M; Jia R; Chen S; Sun K; Yang Q; Wu Y; Chen X; Cheng A
[Ad] Endereço:Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, People's Republic of China liumafengra@163.com chenganchun@vip.163.com.
[Ti] Título:Use of Natural Transformation To Establish an Easy Knockout Method in Riemerella anatipestifer.
[So] Source:Appl Environ Microbiol;83(9), 2017 May 01.
[Is] ISSN:1098-5336
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:is a member of the family and a major causative agent of duck serositis. Little is known about its genetics and pathogenesis. Several bacteria are competent for natural transformation; however, whether is also competent for natural transformation has not been investigated. Here, we showed that strain ATCC 11845 can uptake the chromosomal DNA of strain RA-CH-1 in all growth phases. Subsequently, a natural transformation-based knockout method was established for ATCC 11845. Targeted mutagenesis gave transformation frequencies of ∼10 transformants. Competition assay experiments showed that ATCC 11845 preferentially took up its own DNA rather than heterogeneous DNA, such as DNA. Transformation was less efficient with the shuttle plasmid pLMF03 (transformation frequencies of ∼10 transformants). However, the efficiency of transformation was increased approximately 100-fold using pLMF03 derivatives containing DNA fragments (transformation frequencies of ∼10 transformants). Finally, we found that the RA-CH-1 strain was also naturally transformable, suggesting that natural competence is widely applicable for this species. The findings described here provide important tools for the genetic manipulation of is an important duck pathogen that belongs to the family At least 21 different serotypes have been identified. Genetic diversity has been demonstrated among these serotypes. The genetic and pathogenic mechanisms of remain largely unknown because no genetic tools are available for this bacterium. At present, natural transformation has been found in some bacteria but not in For the first time, we showed that natural transformation occurred in ATCC 11845 and RA-CH-1. Then, we established an easy gene knockout method in based on natural transformation. This information is important for further studies of the genetic diversity and pathogenesis in .
[Mh] Termos MeSH primário: Técnicas de Inativação de Genes/métodos
Genética Microbiana/métodos
Riemerella/genética
Transformação Bacteriana
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1704
[Cu] Atualização por classe:171017
[Lr] Data última revisão:
171017
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170305
[St] Status:MEDLINE


  5 / 7593 MEDLINE  
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[PMID]:28217858
[Au] Autor:Celinska E; Ledesma-Amaro R; Larroude M; Rossignol T; Pauthenier C; Nicaud JM
[Ad] Endereço:Department of Biotechnology and Food Microbiology, Poznan University of Life Sciences, ul. Wojska Polskiego 48, 60-627, Poznan, Poland.
[Ti] Título:Golden Gate Assembly system dedicated to complex pathway manipulation in Yarrowia lipolytica.
[So] Source:Microb Biotechnol;10(2):450-455, 2017 Mar.
[Is] ISSN:1751-7915
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:In this study, we have adopted Golden Gate modular cloning strategy to develop a robust and versatile DNA assembly platform for the nonconventional yeast Yarrowia lipolytica. To this end, a broad set of destination vectors and interchangeable building blocks have been constructed. The DNA modules were assembled on a scaffold of predesigned 4 nt overhangs covering three transcription units (each bearing promoter, gene and terminator), selection marker gene and genomic integration targeting sequences, constituting altogether thirteen elements. Previously validated DNA modules (regulatory elements and selection markers) were adopted as the Golden Gate bricks. The system's operability was demonstrated based on synthetic pathway of carotenoid production. This technology greatly enriches a molecular biology toolbox dedicated to this industrially relevant microorganism enabling fast combinatorial cloning of complex synthetic pathways.
[Mh] Termos MeSH primário: Genética Microbiana/métodos
Engenharia Metabólica/métodos
Redes e Vias Metabólicas
Yarrowia/genética
Yarrowia/metabolismo
[Mh] Termos MeSH secundário: Carotenoides/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
36-88-4 (Carotenoids)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170704
[Lr] Data última revisão:
170704
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170221
[St] Status:MEDLINE
[do] DOI:10.1111/1751-7915.12605


  6 / 7593 MEDLINE  
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[PMID]:28166386
[Au] Autor:Dhakal D; Chung NT; Rayamajhi V; Sohng JK
[Ad] Endereço:Department of Life Science and Biochemical Engineering, Sun Moon University, Asan-si, Republic of Korea.
[Ti] Título:Actinomadura Species: Laboratory Maintenance and Ribosome Engineering.
[So] Source:Curr Protoc Microbiol;44:10G.1.1-10G.1.12, 2017 Feb 06.
[Is] ISSN:1934-8533
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Actinomadura spp. are aerobic, Gram-positive, catalase-positive, non-acid fast, non-motile actinomycetes. Some species of Actinomadura are associated with opportunistic infections in humans. However, many bioactive compounds with pharmaceutical applications can be isolated from various Actinomadura spp. This unit includes general protocols for the laboratory maintenance of Actinomadura spp., including growth in liquid medium, growth on solid agar, long-term storage, and generation of a higher producing strain by ribosome engineering. Actinomadura hibisca P157-2 is used as a prototype for explaining the considerations for efficient laboratory maintenance of Actinomadura spp. © 2017 by John Wiley & Sons, Inc.
[Mh] Termos MeSH primário: Actinobacteria/crescimento & desenvolvimento
Actinobacteria/genética
Técnicas Bacteriológicas/métodos
Genética Microbiana/métodos
Engenharia Metabólica/métodos
Biologia Molecular/métodos
Ribossomos/genética
[Mh] Termos MeSH secundário: Meios de Cultura/química
Preservação Biológica/métodos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Culture Media)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170829
[Lr] Data última revisão:
170829
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170207
[St] Status:MEDLINE
[do] DOI:10.1002/cpmc.22


  7 / 7593 MEDLINE  
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[PMID]:28049774
[Au] Autor:Rosebrock AP
[Ad] Endereço:Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario M5S 3E1, Canada.
[Ti] Título:Synchronization and Arrest of the Budding Yeast Cell Cycle Using Chemical and Genetic Methods.
[So] Source:Cold Spring Harb Protoc;2017(1):pdb.prot088724, 2017 Jan 03.
[Is] ISSN:1559-6095
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The cell cycle of budding yeast can be arrested at specific positions by different genetic and chemical methods. These arrests enable study of cell cycle phase-specific phenotypes that would be missed during examination of asynchronous cultures. Some methods for arrest are reversible, with kinetics that enable release of cells back into a synchronous cycling state. Benefits of chemical and genetic methods include scalability across a large range of culture sizes from a few milliliters to many liters, ease of execution, the absence of specific equipment requirements, and synchronization and release of the entire culture. Of note, cell growth and division are decoupled during arrest and block-release experiments. Cells will continue transcription, translation, and accumulation of protein while arrested. If allowed to reenter the cell cycle, cells will do so as a population of mixed, larger-than-normal cells. Despite this important caveat, many aspects of budding yeast physiology are accessible using these simple chemical and genetic tools. Described here are methods for the block and release of cells in G phase and at the M/G transition using α-factor mating pheromone and the temperature-sensitive cdc15-2 allele, respectively, in addition to methods for arresting the cell cycle in early S phase and at G /M by using hydroxyurea and nocodazole, respectively.
[Mh] Termos MeSH primário: Pontos de Checagem do Ciclo Celular/efeitos dos fármacos
Pontos de Checagem do Ciclo Celular/efeitos da radiação
Divisão Celular/efeitos dos fármacos
Divisão Celular/efeitos da radiação
Genética Microbiana/métodos
Técnicas Microbiológicas/métodos
Saccharomycetales/fisiologia
[Mh] Termos MeSH secundário: Proteínas de Ciclo Celular/genética
Temperatura Alta
Fator de Acasalamento/metabolismo
Proteínas Mutantes/metabolismo
Saccharomycetales/efeitos dos fármacos
Saccharomycetales/crescimento & desenvolvimento
Saccharomycetales/efeitos da radiação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cell Cycle Proteins); 0 (Mutant Proteins); 61194-02-3 (Mating Factor)
[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:170105
[St] Status:MEDLINE
[do] DOI:10.1101/pdb.prot088724


  8 / 7593 MEDLINE  
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[PMID]:28044417
[Au] Autor:Berenguer J; Mencía M; Hidalgo A
[Ad] Endereço:Department of Molecular Biology, Universidad Autónoma de Madrid, Center for Molecular Biology 'Severo-Ochoa' (UAM-CSIC), Nicolás Cabrera 1, Madrid, 28049, Spain.
[Ti] Título:Are in vivo selections on the path to extinction?
[So] Source:Microb Biotechnol;10(1):46-49, 2017 Jan.
[Is] ISSN:1751-7915
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Droplet microfluidics will become a disruptive technology in the field of library screening and replace biological selections if the central dogma of biology and other processes are successfully implemented within microdroplets.
[Mh] Termos MeSH primário: Testes Genéticos
Genética Microbiana/métodos
Técnicas Microbiológicas/métodos
Seleção Genética
[Mh] Termos MeSH secundário: Genótipo
Fenótipo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1706
[Cu] Atualização por classe:170613
[Lr] Data última revisão:
170613
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170104
[St] Status:MEDLINE
[do] DOI:10.1111/1751-7915.12490


  9 / 7593 MEDLINE  
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[PMID]:27811178
[Au] Autor:Krappmann S
[Ad] Endereço:Mikrobiologisches Institut - Klinische Mikrobiologie, Immunologie und Hygiene, Universitätsklinikum Erlangen and Friedrich-Alexander-Universität Erlangen-Nürnberg, Germany sven.krappmann@uk-erlangen.de.
[Ti] Título:CRISPR-Cas9, the new kid on the block of fungal molecular biology.
[So] Source:Med Mycol;55(1):16-23, 2017 Jan 01.
[Is] ISSN:1460-2709
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Research on fungal pathogens with the aim to identify virulence determinants strictly relies on the generation of defined, recombinant strains, a task that is executed by means of a sophisticated molecular biology toolbox. Recent developments in fungal genome engineering have opened a new frontier by implementing the CRISPR-Cas9 technology, based on expression of the Cas9 endonuclease that is loaded by a single guiding RNA (sgRNA) molecule to target a defined site in the recipient genome. This novel approach has been adapted successfully to engineer fungal genomes, among them the one of the human-pathogenic mould Aspergillus fumigatus Implementation of the required components was achieved by various means that differ with respect to expression of the Cas9 enzyme and sgRNA delivery. Validation of CRISPR-Cas9-mediated mutagenesis could be executed by targeting selected candidate genes of A. fumigatus to provide a promising perspective for screening and multiplexing approaches to scrutinize the virulome of this opportunistic fungal pathogen in a comprehensive manner, such as by analyzing genetic polymorphisms or the function of gene families.
[Mh] Termos MeSH primário: Aspergillus fumigatus/genética
Aspergillus fumigatus/fisiologia
Marcação de Genes/métodos
Genética Microbiana/métodos
[Mh] Termos MeSH secundário: Genética Microbiana/tendências
Mutagênese
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170327
[Lr] Data última revisão:
170327
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161105
[St] Status:MEDLINE


  10 / 7593 MEDLINE  
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[PMID]:27730334
[Au] Autor:Zhou C; Shi L; Ye B; Feng H; Zhang J; Zhang R; Yan X
[Ad] Endereço:Department of Microbiology, College of Life Sciences, Key Laboratory for Microbiological Engineering of Agricultural, Environment of Ministry of Agriculture, Nanjing Agricultural University, 6 Tongwei Road, Nanjing, Jiangsu, 210095, People's Republic of China.
[Ti] Título:pheS , an effective host-genotype-independent counter-selectable marker for marker-free chromosome deletion in Bacillus amyloliquefaciens.
[So] Source:Appl Microbiol Biotechnol;101(1):217-227, 2017 Jan.
[Is] ISSN:1432-0614
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:Aside from applications in the production of commercial enzymes and metabolites, Bacillus amyloliquefaciens is also an important group of plant growth-promoting rhizobacteria that supports plant growth and suppresses phytopathogens. A host-genotype-independent counter-selectable marker would enable rapid genetic manipulation and metabolic engineering, accelerating the study of B. amyloliquefaciens and its development as both a microbial cell factory and plant growth-promoting rhizobacteria. Here, a host-genotype-independent counter-selectable marker pheS was constructed through a point mutation of the gene pheS, which encodes the α-subunit of phenylalanyl-tRNA synthetase in Bacillus subtilis strain 168. In the presence of 5 mM p-chloro-phenylalanine, 100 % of B. amyloliquefaciens strain SQR9 cells carrying pheS were killed, whereas the wild-type strain SQR9 showed resistance to p-chloro-phenylalanine. A simple pheS and overlap-PCR-based strategy was developed to create the marker-free deletion of the amyE gene as well as a 37-kb bmy cluster in B. amyloliquefaciens SQR9. The effectiveness of pheS as a counter-selectable marker in B. amyloliquefaciens was further confirmed through the deletion of amyE genes in strains B. amyloliquefaciens FZB42 and NJN-6. In addition, the potential use of pheS in other Bacillus species was preliminarily assessed. The expression of PheS in B. subtilis strain 168 and B. cereus strain ATCC 14579 caused pronounced sensitivity of both hosts to p-chloro-phenylalanine, indicating that pheS could be used as a counter-selectable marker (CSM) in these strains.
[Mh] Termos MeSH primário: Bacillus amyloliquefaciens/genética
Técnicas de Inativação de Genes/métodos
Genética Microbiana/métodos
Fenilalanina-tRNA Ligase/genética
Seleção Genética
[Mh] Termos MeSH secundário: Antibacterianos/toxicidade
Bacillus amyloliquefaciens/fisiologia
Bacillus subtilis/genética
Bacillus subtilis/fisiologia
Fenclonina/toxicidade
Genótipo
Viabilidade Microbiana/efeitos dos fármacos
Proteínas Mutantes/genética
Proteínas Mutantes/metabolismo
Fenilalanina-tRNA Ligase/metabolismo
Mutação Puntual
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anti-Bacterial Agents); 0 (Mutant Proteins); EC 6.1.1.20 (Phenylalanine-tRNA Ligase); R5J7E3L9SP (Fenclonine)
[Em] Mês de entrada:1701
[Cu] Atualização por classe:170620
[Lr] Data última revisão:
170620
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161013
[St] Status:MEDLINE
[do] DOI:10.1007/s00253-016-7906-9



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