Pinto, F.; Pacheco, CC.; Oliveira, P.; Montagud, A.; Landels, A.; Couto, N.; Wright, PC.... (2015). Improving a Synechocystis-based photoautotrophic chassis through systematic genome mapping and validation of neutral sites. DNA Research. 22(6):425-437. https://doi.org/10.1093/dnares/dsv024
Por favor, use este identificador para citar o enlazar este ítem: http://hdl.handle.net/10251/66628
Título:
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Improving a Synechocystis-based photoautotrophic chassis through systematic genome mapping and validation of neutral sites
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Autor:
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Pinto, Filipe
Pacheco, Catarina C.
Oliveira, Paulo
Montagud, Arnau
Landels, Andrew
Couto, Narciso
Wright, Phillip C.
Urchueguía Schölzel, Javier Fermín
Tamagnini, Paula
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Entidad UPV:
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Universitat Politècnica de València. Departamento de Física Aplicada - Departament de Física Aplicada
Universitat Politècnica de València. Instituto Universitario de Matemática Pura y Aplicada - Institut Universitari de Matemàtica Pura i Aplicada
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Fecha difusión:
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Resumen:
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[EN] The use of microorganisms as cell factories frequently requires extensive molecular manipulation.
Therefore, the identification of genomic neutral sites for the stable integration of ectopic DNA is required
to ensure ...[+]
[EN] The use of microorganisms as cell factories frequently requires extensive molecular manipulation.
Therefore, the identification of genomic neutral sites for the stable integration of ectopic DNA is required
to ensure a successful outcome. Herewe describe the genome mapping and validation of five
neutral sites in the chromosome of Synechocystis sp. PCC 6803, foreseeing the use of this cyanobacterium
as a photoautotrophic chassis. To evaluate the neutrality of these loci, insertion/deletion mutants
were produced, and to assess their functionality, a synthetic green fluorescent reporter module
was introduced. The constructed integrative vectors include a BioBrick-compatible multiple cloning
site insulated by transcription terminators, constituting robust cloning interfaces for synthetic biology
approaches. Moreover, Synechocystis mutants (chassis) ready to receive purpose-built synthetic
modules/circuits are also available. This work presents a systematic approach to map and validate
chromosomal neutral sites in cyanobacteria, and that can be extended to other organisms.
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Palabras clave:
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Neutral sites
,
photoautotrophic chassis
,
Synechocystis
,
synthetic biology
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Derechos de uso:
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Reconocimiento - No comercial (by-nc)
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Fuente:
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DNA Research. (issn:
1340-2838
) (eissn:
1756-1663
)
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DOI:
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10.1093/dnares/dsv024
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Editorial:
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Oxford University Press (OUP)
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Versión del editor:
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http://dx.doi.org/10.1093/dnares/dsv024
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Código del Proyecto:
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info:eu-repo/grantAgreement/EC/FP7/308518/EU/Design, construction and demonstration of solar biofuel production using novel (photo)synthetic cell factories/
...[+]
info:eu-repo/grantAgreement/EC/FP7/308518/EU/Design, construction and demonstration of solar biofuel production using novel (photo)synthetic cell factories/
info:eu-repo/grantAgreement/EC/FP6/43340/EU/Engineered modular bacterial hydrogen photo-production of hydrogen/BIOMODULARH2/
info:eu-repo/grantAgreement/UKRI//EP%2FE036252%2F1/GB/ChELSI: Chemical Engineering Life Science Interface/
info:eu-repo/grantAgreement/RCUK/EPSRC/EP/E036252/1/GB/
info:eu-repo/grantAgreement/FCT/SFRH/SFRH/BD/36378/2007/PT/
info:eu-repo/grantAgreement/FCT/SFRH/SFRH/BPD/64095/2009/PT/
info:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBPD%2F74894%2F2010/PT/
info:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBPD%2F64095%2F2009/PT/MODULES AND CIRCUITS FOR H2 PRODUCTION, A SYNTHETIC BIOLOGY APPROACH USING SYNECHOCYSTIS SP. PCC 6803 AS A PHOTOAUTOTROPHIC CHASSIS/
info:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBD%2F36378%2F2007/PT/ENGINEERING OF A CYANOBACTERIUM FOR BIOHYDROGEN PRODUCTION: A NEW COMPUTATIONAL ASSISTED DESIGN OF A PHOTOAUTOTROPHIC CHASSIS/
info:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBPD%2F74894%2F2010/PT/EXPLORING PROTEIN LOCALIZATION AND SECRETION TO DEVELOP CYANOBACTERIA AS ROBUST BIOTECHNOLOGICAL TOOLS/
info:eu-repo/grantAgreement/EC/FP6/043340/EU/Engineered modular bacterial hydrogen photo-production of hydrogen/BIOMODULARH2/
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Agradecimientos:
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This work was supported by the European Commission through the Seventh Framework Programme, FP7-ENERGY-2012-1-2STAGE-308518 (CyanoFactory), from EU FP6-NEST-2005-Path-SYN project BioModularH2 (contract no. 043340) and from ...[+]
This work was supported by the European Commission through the Seventh Framework Programme, FP7-ENERGY-2012-1-2STAGE-308518 (CyanoFactory), from EU FP6-NEST-2005-Path-SYN project BioModularH2 (contract no. 043340) and from National Funds through Fundacao para a Ciencia e a Tecnologia (FCT) (grants SFRH/BD/36378/2007 to F.P., SFRH/BPD/64095/2009 to C.C.P., SFRH/BPD/74894/2010 to P.O.). We also acknowledge the Engineering and Physical Sciences Research Council (EPSRC) for funding (EP/E036252/1) and The University of Sheffield for Scholarship funding. Funding to pay the Open Access publication charges for this article was provided by the European Commission through the Seventh Framework Programme, FP7-ENERGY-2012-1-2STAGE-308518 (CyanoFactory).
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Tipo:
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Artículo
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