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Improving a Synechocystis-based photoautotrophic chassis through systematic genome mapping and validation of neutral sites

Title
Improving a Synechocystis-based photoautotrophic chassis through systematic genome mapping and validation of neutral sites
Type
Article in International Scientific Journal
Year
2015
Authors
Filipe Pinto
(Author)
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Catarina C Pacheco
(Author)
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Paulo Oliveira
(Author)
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Arnau Montagud
(Author)
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Andrew Landels
(Author)
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Narciso Couto
(Author)
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Phillip C Wright
(Author)
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Javier F Urchueguia
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Journal
Title: DNA ResearchImported from Authenticus Search for Journal Publications
Vol. 22
Pages: 425-437
ISSN: 1340-2838
Scientific classification
FOS: Natural sciences > Earth and related Environmental sciences
Other information
Authenticus ID: P-00J-YXF
Abstract (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. Here we 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.
Language: English
Type (Professor's evaluation): Scientific
No. of pages: 13
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