Título:	Synthetic biology toolbox for nitrogen-fixing soil microbes.
Autoria:	VENKATARAMAN, M. YÑIGEZ-GUTIERREZ, A. INFANTE, V. MACINTYRE, A. FERNANDES JUNIOR, P. I. ANÉ, J.-M. PFLEGER, B.
Afiliação:	MAYA VENKATARAMAN AUDREY YÑIGEZ-GUTIERREZ VALENTINA INFANTE APRIL MACINTYRE PAULO IVAN FERNANDES JUNIOR, CPATSA JEAN-MICHEL ANÉ BRIAN PFLEGER.
Ano de publicação:	2023
Referência:	ACS Synthetic Biology, v. 12, n. 12, p. 3623-3634, 2023.
Conteúdo:	The 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.
Thesagro:	Solo Nitrogenase
NAL Thesaurus:	Synthetic biology Genome
Palavras-chave:	CRISPR Cas9 Caixa de ferramentas Biologia sintética Edição de genoma
Digital Object Identifier:	https://doi.org/10.1021/acssynbio.3c00414
Tipo do material:	Artigo de periódico
Acesso:	openAccess
Aparece nas coleções:	Artigo em periódico indexado (CPATSA)