A highly precise and portable genome engineering method allows comparison of mutational effects across bacterial species (NDA@SZTAKI, Nemzeti Digitális Adattár, National Digital Data Archive)

A highly precise and portable genome engineering method allows comparison of mutational effects across bacterial species

Metaadatok

Tartalom:	http://real.mtak.hu/48959/
Archívum:	MTA Könyvtár
Gyûjtemény:	Status = Published Type = Article
Cím:	A highly precise and portable genome engineering method allows comparison of mutational effects across bacterial species
Létrehozó:	Nyerges, Ãkos CsÃ¶rgÅ‘, BÃ¡lint Nagy, IstvÃ¡n BÃ¡lint, BalÃ¡zs Bihari, PÃ©ter LÃ¡zÃ¡r, ViktÃ³ria Apjok, GÃ¡bor Umenhoffer, Kinga Bogos, BalÃ¡zs PÃ³sfai, GyÃ¶rgy PÃ¡l, Csaba
Kiadó:	National Academy of Sciences of Armenia
Dátum:	2016
Téma:	QH301 Biology / biolÃ³gia
Tartalmi leírás:	Currently available tools for multiplex bacterial genome engineering are optimized for a few laboratory model strains, demand extensive prior modification of the host strain, and lead to the accumulation of numerous off-target modifications. Building on prior development of multiplex automated genome engineering (MAGE), our work addresses these problems in a single framework. Using a dominant-negative mutant protein of the methyl-directed mismatch repair (MMR) system, we achieved a transient suppression of DNA repair in Escherichia coli, which is necessary for efficient oligonucleotide integration. By integrating all necessary components into a broad-host vector, we developed a new workflow we term pORTMAGE. It allows efficient modification of multiple loci, without any observable off-target mutagenesis and prior modification of the host genome. Because of the conserved nature of the bacterial MMR system, pORTMAGE simultaneously allows genome editing and mutant library generation in other biotechnologically and clinically relevant bacterial species. Finally, we applied pORTMAGE to study a set of antibiotic resistance-conferring mutations in Salmonella enterica and E. coli. Despite over 100 million y of divergence between the two species, mutational effects remained generally conserved. In sum, a single transformation of a pORTMAGE plasmid allows bacterial species of interest to become an efficient host for genome engineering. These advances pave the way toward biotechnological and therapeutic applications. Finally, pORTMAGE allows systematic comparison of mutational effects and epistasis across a wide range of bacterial species.
Nyelv:	angol
Típus:	Article PeerReviewed info:eu-repo/semantics/article
Formátum:	text
Azonosító:	http://real.mtak.hu/48959/1/NyergesPNAS.pdf Nyerges, Ãkos and CsÃ¶rgÅ‘, BÃ¡lint and Nagy, IstvÃ¡n and BÃ¡lint, BalÃ¡zs and Bihari, PÃ©ter and LÃ¡zÃ¡r, ViktÃ³ria and Apjok, GÃ¡bor and Umenhoffer, Kinga and Bogos, BalÃ¡zs and PÃ³sfai, GyÃ¶rgy and PÃ¡l, Csaba (2016) A highly precise and portable genome engineering method allows comparison of mutational effects across bacterial species. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 113 (9). pp. 2502-2507. ISSN 0027-8424
Kapcsolat:	http://real.mtak.hu/48959/ http://doi.org/10.1073/pnas.1520040113 MTMT:3038670; doi:10.1073/pnas.1520040113