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Optimization of electroporation method and promoter evaluation for type-1 methanotroph, Methylotuvimicrobium alcaliphilum.
Goswami, Shubhasish; Singer, Steven W; Simmons, Blake A; Awasthi, Deepika.
Afiliación
  • Goswami S; Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, United States.
  • Singer SW; Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, United States.
  • Simmons BA; Joint BioEnergy Institute, Emeryville, CA, United States.
  • Awasthi D; Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, United States.
Front Bioeng Biotechnol ; 12: 1412410, 2024.
Article en En | MEDLINE | ID: mdl-38812915
ABSTRACT
Methanotrophic bacteria are promising hosts for methane bioconversion to biochemicals or bioproducts. However, due to limitations associated with long genetic manipulation timelines and, lack of choice in genetic tools required for strain engineering, methanotrophs are currently not employed for bioconversion technologies. In this study, a rapid and reproducible electroporation protocol is developed for type 1 methanotroph, Methylotuvimicrobium alcaliphilum using common laboratory solutions, analyzing optimal electroshock voltages and post-shock cell recovery time. Successful reproducibility of the developed method was achieved when different replicative plasmids were assessed on lab adapted vs. wild-type M. alcaliphilum strains (DASS vs. DSM19304). Overall, a ∼ 3-fold decrease in time is reported with use of electroporation protocol developed here, compared to conjugation, which is the traditionally employed approach. Additionally, an inducible (3-methyl benzoate) and a constitutive (sucrose phosphate synthase) promoter is characterized for their strength in driving gene expression.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Front Bioeng Biotechnol Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Suiza
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Front Bioeng Biotechnol Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Suiza