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dc.contributor.authorVENKATARAMAN, M.
dc.contributor.authorYÑIGEZ-GUTIERREZ, A.
dc.contributor.authorINFANTE, V.
dc.contributor.authorMACINTYRE, A.
dc.contributor.authorFERNANDES JUNIOR, P. I.
dc.contributor.authorANÉ, J.-M.
dc.contributor.authorPFLEGER, B.
dc.date.accessioned2023-12-18T13:32:30Z-
dc.date.available2023-12-18T13:32:30Z-
dc.date.created2023-12-18
dc.date.issued2023
dc.identifier.citationACS Synthetic Biology, v. 12, n. 12, p. 3623-3634, 2023.
dc.identifier.urihttp://www.alice.cnptia.embrapa.br/alice/handle/doc/1159849-
dc.descriptionThe soil environment adjacent to plant roots, termed the rhizosphere, is home to a wide variety of microorganisms that can significantly affect the physiology of nearby plants. Microbes in the rhizosphere can provide nutrients, secrete signaling compounds, and inhibit pathogens. These processes could be manipulated with synthetic biology to enhance the agricultural performance of crops grown for food, energy, or environmental remediation, if methods can be implemented in these nonmodel microbes. A common first step for domesticating nonmodel organisms is the development of a set of genetic engineering tools, termed a synthetic biology toolbox. A toolbox comprises transformation protocols, replicating vectors, genome engineering (e.g., CRISPR/Cas9), constitutive and inducible promoter systems, and other gene expression control elements. This work validated synthetic biology toolboxes in three nitrogen-fixing soil bacteria: Azotobacter vinelandii, Stutzerimonas stutzeri (Pseudomonas stutzeri), and a new isolate of Klebsiella variicola. All three organisms were amenable to transformation and reporter protein expression, with several functional inducible systems available for each organism. S. stutzeri and K. variicola showed more reliable plasmid-based expression, resulting in successful Cas9 recombineering to create scarless deletions and insertions. Using these tools, we generated mutants with inducible nitrogenase activity and introduced heterologous genes to produce resorcinol products with relevant biological activity in the rhizosphere.
dc.language.isoeng
dc.rightsopenAccess
dc.subjectCRISPR
dc.subjectCas9
dc.subjectCaixa de ferramentas
dc.subjectBiologia sintética
dc.subjectEdição de genoma
dc.titleSynthetic biology toolbox for nitrogen-fixing soil microbes.
dc.typeArtigo de periódico
dc.subject.thesagroSolo
dc.subject.thesagroNitrogenase
dc.subject.nalthesaurusSynthetic biology
dc.subject.nalthesaurusGenome
riaa.ainfo.id1159849
riaa.ainfo.lastupdate2023-12-18
dc.identifier.doihttps://doi.org/10.1021/acssynbio.3c00414
dc.contributor.institutionMAYA VENKATARAMAN
dc.contributor.institutionAUDREY YÑIGEZ-GUTIERREZeng
dc.contributor.institutionVALENTINA INFANTEeng
dc.contributor.institutionAPRIL MACINTYREeng
dc.contributor.institutionPAULO IVAN FERNANDES JUNIOR, CPATSAeng
dc.contributor.institutionJEAN-MICHEL ANÉeng
dc.contributor.institutionBRIAN PFLEGER.eng
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