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

• Tipo: Artigo em Revista Científica Internacional
• Data: 2015

Autores

Filipe Pinto (Autor) Outra; Catarina C Pacheco (Autor) Outra; Paulo Oliveira (Autor) Outra; Arnau Montagud (Autor) Outra; Andrew Landels (Autor) Outra; Narciso Couto (Autor) Outra; Phillip C Wright (Autor) Outra; Javier F Urchueguia (Autor) Outra; Paula Tamagnini (Autor) FCUP

Revista

Título: DNA Research

Vol. 22

Páginas: 425-437

ISSN: 1340-2838

Editora: Oxford University Press

Indexação

ISI Web of Knowledge - 0 Citações

Scopus - 44 Citações

Classificação Científica

FOS: Ciências exactas e naturais > Ciências da terra e ciências do ambiente

Outras Informações

ID Authenticus: P-00J-YXF

DOI: 10.1093/dnares/dsv024

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.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 13