| Full text | |
| Author(s): |
Westmann, Caua A.
[1]
;
Alves, Luana de Fatima
[2, 3]
;
Silva-Rocha, Rafael
[1]
;
Guazzaroni, Maria-Eugenia
[2]
Total Authors: 4
|
| Affiliation: | [1] Univ Sao Paulo, Dept Cellular & Mol Biol, FMRP, Ribeirao Preto - Brazil
[2] Univ Sao Paulo, Dept Biol, FFCLRP, Ribeirao Preto - Brazil
[3] Univ Sao Paulo, Dept Biochem, FMRP, Ribeirao Preto - Brazil
Total Affiliations: 3
|
| Document type: | Journal article |
| Source: | FRONTIERS IN MICROBIOLOGY; v. 9, JUN 20 2018. |
| Web of Science Citations: | 5 |
| Abstract | |
Although functional metagenomics has been widely employed for the discovery of genes relevant to biotechnology and biomedicine, its potential for assessing the diversity of transcriptional regulatory elements of microbial communities has remained poorly explored. Here, we experimentally mined novel constitutive promoter sequences in metagenomic libraries by combining a bi-directional reporter vector, high-throughput fluorescence assays and predictive computational methods. Through the expression profiling of fluorescent clones from two independent soil sample libraries, we have analyzed the regulatory dynamics of 260 clones with candidate promoters as a set of active metagenomic promoters in the host Escherichia coli. Through an in-depth analysis of selected clones, we were able to further explore the architecture of metagenomic fragments and to report the presence of multiple promoters per fragment with a dominant promoter driving the expression profile. These approaches resulted in the identification of 33 novel active promoters from metagenomic DNA originated from very diverse phylogenetic groups. The in silico and in vivo analysis of these individual promoters allowed the generation of a constitutive promoter consensus for exogenous sequences recognizable by E. coli in metagenomic studies. The results presented here demonstrates the potential of functional metagenomics for exploring environmental bacterial communities as a source of novel regulatory genetic parts to expand the toolbox for microbial engineering. (AU) | |
| FAPESP's process: | 16/06323-4 - New tools for improving metagenomic cellulase screenings |
| Grantee: | Luana de Fátima Alves |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| FAPESP's process: | 15/04309-1 - Novel approaches to improve functional screening of biocatalysts in metagenomic libraries |
| Grantee: | María Eugenia Guazzaroni |
| Support Opportunities: | Research Grants - Young Investigators Grants |
| FAPESP's process: | 16/05472-6 - Synthetic biology approaches to engineer/mine novel regulatory elements in bacteria |
| Grantee: | Cauã Antunes Westmann |
| Support Opportunities: | Scholarships in Brazil - Master |