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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Expanding the Toolbox of Broad Host-Range Transcriptional Terminators for Proteobacteria through Metagenomics

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Amarelle, Vanesa [1, 2] ; Sanches-Medeiros, Ananda [3] ; Silva-Rocha, Rafael [3] ; Guazzaroni, Maria-Eugenia [1]
Total Authors: 4
[1] Univ Sao Paulo, FFCLRP, BR-14049901 Ribeirao Preto, SP - Brazil
[2] Biol Res Inst Clemente Estable, Dept Microbial Biochem & Genom, Montevideo 11600 - Uruguay
[3] Univ Sao Paulo, FMRP, BR-14049901 Ribeirao Preto, SP - Brazil
Total Affiliations: 3
Document type: Journal article
Source: ACS SYNTHETIC BIOLOGY; v. 8, n. 4, p. 647-654, APR 2019.
Web of Science Citations: 1

As the field of synthetic biology moves toward the utilization of novel bacterial chassis, there is a growing need for biological parts with enhanced performance in a wide number of hosts. Is not unusual that biological parts (such as promoters and terminators), initially characterized in the model bacterium Escherichia coli, do not perform well when implemented in alternative hosts, such as Pseudomonas, therefore limiting the construction of synthetic circuits in industrially relevant bacteria, for instance Pseudomonas putida. In order to address this limitation, we present here the mining of transcriptional terminators through functional metagenomics to identify novel parts with broad host-range activity. Using a GFP-based terminator trap strategy and a broad host-range plasmid, we identified 20 clones with potential terminator activity in P. putida. Further characterization allowed the identification of 4 unique sequences ranging from 58 to 181 bp long that efficiently terminate transcription in P. putida, E. coli, Burkholderia phymatum, and two Pseudomonas strains isolated from Antarctica. Therefore, this work presents a new set of biological parts useful for the engineering of synthetic circuits in Proteobacteria. (AU)

FAPESP's process: 18/04810-0 - Deconstructing complexity in the regulatory network for biofilm formation in gram-negative bacteria
Grantee:Ananda Sanches Medeiros
Support type: Scholarships in Brazil - Doctorate (Direct)
FAPESP's process: 15/04309-1 - Novel approaches to improve functional screening of biocatalysts in metagenomic libraries
Grantee:María Eugenia Guazzaroni
Support type: Research Grants - Young Investigators Grants