| RRC ID |
53714
|
| 著者 |
Pinto F, Pacheco CC, Oliveira P, Montagud A, Landels A, Couto N, Wright PC, Urchueguía JF, Tamagnini P.
|
| タイトル |
Improving a Synechocystis-based photoautotrophic chassis through systematic genome mapping and validation of neutral sites.
|
| ジャーナル |
DNA Res
|
| Abstract |
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.
|
| 巻・号 |
22(6)
|
| ページ |
425-37
|
| 公開日 |
2015-12-1
|
| DOI |
10.1093/dnares/dsv024
|
| PII |
dsv024
|
| PMID |
26490728
|
| PMC |
PMC4675711
|
| MeSH |
Autotrophic Processes
Chromosome Mapping
Cyanobacteria / genetics*
Cyanobacteria / growth & development
Green Fluorescent Proteins
Mutation
Synechocystis / genetics*
Synthetic Biology / methods
|
| IF |
4.009
|
| 引用数 |
19
|
| リソース情報 |
| 原核生物(大腸菌) |
|
